Angelica polysaccharide attenuates LPS-induced inflammation response of primary dairy cow claw dermal cells via NF-κB and MAPK signaling pathways.

BACKGROUND: Laminitis, an inflammation of the claw laminae, is one of the major causes of bovine lameness, which can lead to enormous economic losses and animal welfare problems in dairy farms. Angelica polysaccharide (AP) is proved to possess anti-inflammatory properties. But the role of AP on inflammatory response of the claw dermal cells has not been reported. The aim of this study was to investigate the anti-inflammatory effects of AP on lipopolysaccharide (LPS)-induced primary claw dermal cells of dairy cow and clarify the potential mechanisms. In the current research, the primary claw dermal cells were exposed to gradient concentrations of AP (10, 50, 100 µg/mL) in the presence of 10 µg/mL LPS. The levels of cytokines and nitric oxide (NO) were detected with ELISA and Griess colorimetric method. The mRNA expressions of TLR4, MyD88 and chemokines were measured with qPCR. The activation of NF-κB and MAPK signaling pathways was detected with western blotting. RESULTS: The results indicated that AP reduced the production of inflammatory mediators (TNF-α, IL-1ß, IL-6 and NO), downregulated the mRNA expression of TLR4, MyD88 and some pro-inflammatory chemokines (CCL2, CCL20, CXCL2, CXCL8, CXCL10), and suppressed the NF-κB and MAPK signaling pathways evidenced by inhibition of the phosphorylation of IκBα, p65 and ERK, JNK, p38. CONCLUSIONS: Our results demonstrated that AP may exert its anti-inflammatory effects on claw dermal cells of dairy cow by regulating the NF-κB and MAPK signaling pathways.

Effect of Penetration Enhancers on Toenail Delivery of Efinaconazole from Hydroalcoholic Preparations.

The incorporation of permeation enhancers in topical preparations has been recognized as a simple and valuable approach to improve the penetration of antifungal agents into toenails. In this study, to improve the toenail delivery of efinaconazole (EFN), a triazole derivative for onychomycosis treatment, topical solutions containing different penetration enhancers were designed, and the permeation profiles were evaluated using bovine hoof models. In an in vitro permeation study in a Franz diffusion cell, hydroalcoholic solutions (HSs) containing lipophilic enhancers, particularly prepared with propylene glycol dicaprylocaprate (Labrafac PG), had 41% higher penetration than the HS base. Moreover, the combination of hydroxypropyl-ß-cyclodextrin with Labrafac PG further facilitated the penetration of EFN across the hoof membrane. In addition, this novel topical solution prepared with both lipophilic and hydrophilic enhancers was physicochemically stable, with no drug degradation under ambient conditions (25 °C, for 10 months). Therefore, this HS system can be a promising tool for enhancing the toenail permeability and therapeutic efficacy of EFN.

A novel technique to evaluate nail softening effects of different urea formulations.

Urea has been incorporated into several topical ungual formulations to hydrate and soften the nail plate. In this study, we employed various characterization techniques (visual observation, scanning electron microscopy, measurement of thickness, transonychial water loss, nail electrical resistance, and mechanical study) to investigate the effect of urea concentration on the hydration of bovine hoof membranes - an in vitro model of infected human nails. We obtained inconsistent results in the thickness, transonychial water loss, nail electrical resistance, and scanning electron microscopy studies. In the mechanical study using a modified Texture Analyzer method, we reported an inverse and linear correlation between urea concentrations in the formulations and the force required to puncture the treated membrane (R2 = 0.9582, n ≥ 8). As the urea concentration decreased from 4x to 2x, 1x, and 0x % w/w, the puncture force increased significantly from 0.47 ± 0.07 to 0.77 ± 0.07, 0.91 ± 0.09, and 1.33 ± 0.26 N, respectively (p < 0.05). Thus, urea provided a positive softening effect on the membranes and the puncture force could indicate the urea level in topical formulations. In this study, we provided a novel, efficient, and reliable tool to evaluate the hydration level and physical properties of bovine hoof membranes.

Potential of Naftifine Application for Transungual Delivery.

Naftifine is used to treat fungal skin infections as it inhibits dermatophytes, which are the cause of onychomycosis. However, naftifine's ability to permeate the human nail barrier has not been investigated, thus, the antimycotic potential is not clearly established. This work aims to evaluate the effect of penetration enhancing factors on the accumulation of naftifine hydrochloride through human nail clippings. Naftifine polymeric nail lacquers with Eudragit RL100 were developed as a suitable delivery system. Low penetration of naftifine into nail has been determined as less than 10% of applied drug dose accumulated in the nail layers. Incorporation of thioglycolic acid into formulations resulted in increased accumulation of antifungal agent in the nail layers by 100% compared with a control group. Salicylic acid did not effect naftifine accumulation in the human nail. The permeation of naftifine through the nail increased by threefold when the thioglycolic acid-containing formulation was applied and the nail was pretreated with a fractional CO2 laser. Structural changes of the nail barrier, induced by fractional CO2 laser, were visualized by microscopy. The results suggest, that naftifine nail penetration could be significantly increased when physical and chemical enhancing factors are applied.

Ciclopirox and Efinaconazole Transungual Permeation, Antifungal Activity, and Proficiency To Induce Resistance in Trichophyton rubrum.

Onychomycosis is a nail fungal infection, mostly caused by dermatophytes. The treatment efficacy is impaired by difficulties of reaching effective drug levels at the site of infection; frequent relapses occur after cessation of antifungal therapy. The aim of the study was to compare two commercial products containing ciclopirox or efinaconazole for antimycotic activity and antifungal drug resistance. A study of permeation and penetration through bovine hoof membranes, as a nail model, was performed to evaluate the antimycotic activity of permeates against clinical isolates of selected fungi, and the frequency of spontaneous in vitroTrichophyton rubrum-resistant strains was assessed by broth microdilution assays. The results suggest that ciclopirox creates a depot in the nail, leading to a gradual release of the drug over time with action on both the nail plate and bed. Conversely, efinaconazole, mildly interacting with nail keratin, mainly exerts its antifungal activity in the nail bed. However, in the case of T. rubrum, the antifungal activities of the drugs in the nail plate seem comparable. Finally, efinaconazole showed a potential for induction of resistance in T. rubrum, which may limit its efficacy over time. Ciclopirox did not show any potential to induce resistance in T. rubrum and appears endowed with a more complete activity than efinaconazole in the management of onychomycosis as the nail keratin is a substrate for the growth of fungal cells, and the availability of drug in large concentration just in the nail bed may not be sufficient to guarantee the complete eradication of pathogens.

Effects of silymarin on p65 NF-κB, p38 MAPK and CYP450 in LPS-induced hoof dermal inflammatory cells of dairy cows.

BACKGROUND: Laminitis is considered as one of the most important causes of hoof lameness in dairy cows, which can lead to enormous economic losses. However, the etiology and pathogenesis of laminitis have not been clarified yet. Besides, it is of great significant to find alternative herbs for the prevention and treatment of dairy hooves to avoid the antibiotic abuse. In this study, the primary hoof dermal cells of dairy cows were isolated, the inflammatory model was induced by LPS, and treated with silymarin to find whether silymarin has protective effect on the inflammatory dermal cells. The viability of dermal cells, the levels of IL-1ß and TNF-α, the degree of p65 NF-κB and p38 MAPK phosphorylation, the expressions of CYP3A4 and CYP1A1 were measured. RESULTS: Hoof dermal cells of dairy cows were successfully isolated and cultured by tissue adherent culture method. Certain concentrations of LPS can increase the levels of IL-1ß and TNF-α, promote the phosphorylation of p65 NF-κB and p38 MAPK, and inhibit the mRNA expressions of CYP3A4 and CYP1A1. The optimal concentration for LPS to establish a hoof dermal cells inflammatory model was 10 µg/mL. Certain concentrations of silymarin can markedly decrease the secretions of IL-1ß and TNF-α, inhibit the phosphorylation of p65 NF-κB and p38 MAPK, and promote the mRNA expressions of CYP3A4 and CYP1A1 in LPS-induced dermal inflammatory model. CONCLUSIONS: LPS can be used for inducing the hoof dermal cells inflammatory model of dairy cows. Silymarin has protective effects on the LPS-induced inflammatory model.

Use of topical local anesthetics to control pain during treatment of hoof lesions in dairy cows.

Hoof pathologies in dairy cows have a major effect on both production and animal welfare. Trimming of excess or diseased hoof tissue is essential for the treatment of many of these conditions. Trimming hoof lesions can cause severe pain, resulting in adverse behavioral responses with risk for animal and human safety. Interventions are usually carried out by nonveterinary technicians in the absence of pain management training. Pain control during trimming is not only an ethical obligation but also allows for better manipulation and more meticulous treatment. The aim of this study was to test the efficacy of Tri-Solfen (Bayer Australia Ltd., Pymble, NSW, Australia), a combination of local anesthetics in a topical gel form, containing lidocaine, bupivacaine, adrenaline, and cetrimide, for the treatment of pain associated with trimming of hoof lesions. Sixty-two Holstein-Frisian cows were selected for trimming at the drying-off period and were visually scored for lameness before entering the chute. After diagnosis of the hoof lesion but before deep trimming was initiated, each animal was randomly distributed to 2 groups: C, usual trimming with no pain control, and T, trimming with a local anesthetic formulation being applied immediately after live corium was exposed. During curative trimming, behavior observation was conducted by 2 observers blind to treatment. In 27 cows, algometry measurements were performed before and after the procedure to assess animal reaction to pressure. Lameness scoring was again performed as the cow left the chute. Nonparametric tests and ANOVA were performed. Results showed that use of the topical anesthetic formulation significantly reduced reaction to trimming and lameness score after trimming when compared with nontreated animals. Algometry values showed increased pressure threshold after application of topical anesthetics. This study suggests that the use of topical local anesthesia with lidocaine and bupivacaine helps reduce pain associated with corrective trimming of severe hoof lesions, enhancing animal welfare and potentially ensuring safety of trimmers.

Comparison of effects of routine topical treatments in the milking parlor on digital dermatitis lesions.

Digital dermatitis (DD), an infectious bacterial disease affecting the feet of dairy cattle, can cause lameness and decrease milk production, fertility, and animal welfare. Current DD treatment typically involves routine hoof trimming and topical antibiotics. Several nonantibiotic commercial topical products are used for controlling DD lesions; however, there is limited or no evidence regarding their effectiveness. The objectives of this study were to evaluate 2 commercially available topical applications on their ability to (1) clinically cure active DD lesions to nonactive lesions and (2) prevent recurrence of active DD lesions. Ten farms were visited weekly. In the milking parlor, the hind feet of lactating cattle were cleaned and scored (M-stage scoring system). Cattle with DD lesions at the first visit were randomly allocated to 1 of 4 treatment groups: positive control (tetracycline solution), HealMax (AgroChem Inc., Saratoga Springs, NY), HoofSol (Diamond Hoof Care Ltd., Intracare BV, Veghel, the Netherlands), and a negative control (saline). All products were applied to lesions using a spray bottle. Tetracycline, HealMax, and HoofSol had a higher probability of clinical cure for active lesions compared with saline 1 wk after the first treatment (wk 1), with 69, 52, and 79% clinical cure of active lesions, respectively, compared with 34% with saline. At wk 7, the probability of clinical cure for active lesions was 10, 33, 31, and 45% of lesions treated weekly with saline, tetracycline, HealMax, and HoofSol, respectively (no difference among treatments). The substantial clinical cure with saline highlighted the potential importance of cleaning feet. In wk 1, treatment with saline, tetracycline, HealMax, and HoofSol resulted in a probability of recurrence of active DD lesions of 9, 11, 11, and 8%, respectively, with no product being superior to saline. After 7 wk, the probability of recurrence of active lesions was 5, 7, 6, and 6% for saline, tetracycline, HealMax, and HoofSol respectively, with no difference among groups in wk 7. These results provide alternatives to antibiotics for treatment of DD lesions and highlight the potential importance of cleaning feet in the milking parlor.

Influence of a Combination of Chemical Enhancers and Iontophoresis on In Vitro Transungual Permeation of Nystatin.

To promote transungual permeation of nystatin (NYST), molecule with high molecular weight, no water-soluble, amphoteric by iontophoresis. The synergic effect of the combination of cetylpyridinium chloride, CPC, or polyoxyethylene (20) sorbitan monooleate, TW80, and iontophoresis was investigated. In vitro permeation experiments were carried out through bovine hoof slices using vertical diffusion cells. A low current density (0.2 mA/cm2) was applied by introducing Ag/AgCl electrodes in the donor (anode) and receptor (cathode) chambers. The donor phase consisted of a solution, a suspension, or gel-type vehicles containing NYST and surfactants in pH 5.6 HEPES buffer. The addition of CPC to NYST suspension (SOSP) produced a fivefold increase on the permeability of the bovine hoof membrane to the drug. The application of anodal iontophoresis further improved NYST flux. Conversely, NYST transungual permeation was not influenced by TW80 either in the passive diffusion or iontophoretic flux. Furthermore, the iontophoretic treatment does not appear to induce irreversible alterations to the hoof bovine membranes. The present work demonstrated the efficacy of iontophoresis as a treatment for different nail pathologies with large molecules very slightly soluble in water without irreversibly affecting the nail structure. A synergistic effect between CPC and iontophoresis was observed.

Evaluation of topical antifungal products in an in vitro onychomycosis model.

Many topical commercial products are currently available for the treatment of onychomycosis. However, limited data are available concerning their antifungal activity. Using an in vitro onychomycosis model, the daily application of seven nail formulations was compared to the antifungal reference drug amorolfine (Loceryl(®) ) and evaluated for inhibitory activity against Trichophyton mentagrophytes using an agar diffusion test. Of all commercial nail formulations, only Excilor(®) and Nailner(®) demonstrated inhibitory activity, which was much lower compared to the daily application of Loceryl(®) . However, Excilor(®) showed similar efficacy compared to the conventional weekly application of Loceryl(®) . These results suggest a role for organic acids in the antifungal effect of Excilor(®) (acetic acid, ethyl lactate) and Nailner(®) (lactic acid, citric acid, ethyl lactate) as all tested formulations without organic acids were inactive.

Effectiveness of different regimens of a collective topical treatment using a solution of copper and zinc chelates in the cure of digital dermatitis in dairy farms under field conditions.

A controlled field trial was conducted to evaluate in dairy cattle the benefit provided by different regimens of a collective topical treatment using a solution of copper and zinc chelates to cure digital dermatitis (DD) compared with individual treatment alone, and further to investigate factors that could explain variations in the clinical cure of DD lesions over 6 mo. The study was conducted between November 2009 and October 2010 and involved 4,677 dairy cows from 52 French dairy farms on which DD was endemic. The farms were quasi-randomly allocated to 1 of 4 treatment regimens for 6 mo: no collective treatment (control), walk-through footbath during 4 consecutive milkings every 4 wk (FB/4W) or every 2 wk (FB/2W) and collective spraying during 2 milkings every 2 wk (CS/2W). For ethical and welfare reasons, all farmers also had to treat all detected active DD lesions with individual topical spraying of oxytetracycline. Digital dermatitis and leg hygiene were scored on all lactating cows during milking 7 times every 4 wk by 14 trained investigators. During these farm visits, data related to farm management were also collected. The curative effectiveness of collective treatments was assessed through a Cox survival frailty model as the probability of cure of an active DD lesion during at least 2 consecutive visits. The model was adjusted for farm and cow risk factors as well as initial DD prevalence. Monthly DD cure rates were 58, 55, 76, and 76% in the control, FB/4W, FB/2W, and CS/2W regimens, respectively. The spontaneous monthly cure rate for untreated active DD lesions was 61%. Hazard of cure of DD was increased by 1.28 and 1.41 when walk-through footbath and collective spraying, respectively, were applied over 2 d every 2 wk compared with the control regimen. Applying a walk-through footbath 2 d every 4 wk was not sufficient to improve the cure of DD compared with individual treatments alone. Three main factors were identified as speeding DD healing: cleanliness of the feet, initial small size of the DD lesion, and additional individual topical treatment. Grazing tended to speed DD healing. These results highlight the need of combining several control measures, including individual and collective topical treatments, and improving foot hygiene and the early detection of DD lesions to ensure a high cure rate and rapid curing of digital dermatitis on endemically affected farms.

Effectiveness of different footbathing frequencies using copper sulfate in the control of digital dermatitis in dairy cows.

Two experiments were conducted to test the effectiveness of different footbathing frequencies using CuSO4 in the control of digital dermatitis (DD) in groups of dairy cows with low and high DD prevalence in an endemically infected herd. During the study, groups of cows walked through allocated footbath solutions after milking on 4 consecutive milkings each week, 2 wk, or monthly, depending on treatment. The footbath solutions were changed either after 200 cows had passed through the footbath or within 24 h. All cows were scored weekly during milking for DD lesion stage on the hind feet using a 5-point nominal scale. A transition grade was assigned based on whether the DD lesions improved (1) or deteriorated or did not improve (0) from week to week, and was averaged for all cows in the group. Furthermore, from the longitudinal study data, all transitions between different DD lesion stages between the different time points were used in a discrete, first-order Markov chain (state transition matrix) model. In experiment 1, 70 Holstein-Friesian cows with DD lesions present on at least one of their hind feet were allocated to 1 of 2 footbath treatments for 14 wk: (1) 5% CuSO4 each week or (2) 5% CuSO4 every 2 wk. At the end of the study, no active DD lesions were observed in cows in either treatment group, but significantly more cows had no DD lesions (0.48 ± 0.097 vs. 0.24 ± 0.094) and fewer cows had lesions in the healing stage (0.52 ± 0.104 vs. 0.77 ± 0.090) for the weekly compared with the 2-wk footbathing regimen. The number of active DD lesions that the transition matrix model predicted over time was 3 and 8% for weekly and every 2 wk footbathing treatments, respectively. In experiment 2, 64 cows with no DD lesions on either of their hind feet were allocated to 1 of 2 footbath treatment regimens for 14 wk: (1) 5% CuSO4 every 2 wk or (2) 5% CuSO4 each month. At the end of the trial, significantly more cows had no DD (0.80 ± 0.088 vs. 0.65 ± 0.102) and fewer cows had DD lesions in the healing stage (0.20 ± 0.088 vs. 0.35 ± 0.102) in the every 2 wk regimen than in the monthly footbathing regimen. The number of active DD lesions that the transition matrix model predicted over time was low for both footbathing interventions (1 and 4%, respectively). Increasing the interval between footbaths does not appear the most appropriate mechanism for reducing CuSO4 usage.

Validation of bovine hoof slices as a model for infected human toenails: in vitro ciclopirox transungual permeation.

BACKGROUND: Topical therapy has recently been proposed for treating onychomycosis and other nail disturbances. However, the clinical outcome may be limited by the difficulty of active ingredients effectively penetrating the nail plate. Bovine hoof membranes have been widely used to predict in vitro efficacy of drug products in nail diseases. Many studies have compared bovine hooves with human healthy nails, considering the difference between healthy and unhealthy nails to be negligible. OBJECTIVES: To validate bovine hoof slices as a model for human unhealthy nails by investigating the transungual permeation/retention of ciclopirox (CPX) through bovine hoof slices and excised infected human toenails after application of a new film-forming formulation (P-3051). To investigate the ability of CPX to achieve fungicidal concentrations in and through infected toenails. METHODS: A new experimental technique based on a permeation unit allowed analysis by high-performance liquid chromatography of the amounts of CPX permeating through and retained in the membranes. The efficacy index was evaluated as follows: amount of permeated CPX/Trichophyton rubrum minimum inhibiting concentration. RESULTS: Extrapolated CPX flux through bovine hoof slices was about 14-fold higher than through infected human toenails, the difference being mainly due to the fourfold higher thickness of the toenails. In toenails, the CPX efficacy index for T. rubrum was positive (>1·0) soon after P-3051 application. CONCLUSIONS: This study confirms the validity of bovine hoof slices as a model for infected human nails, and suggests a substantial equivalence between the two models. Following P-3051 application, CPX reaches fungicidal concentrations in and through human infected toenails.

Effect of feeding biotin on milk production and hoof health in lactating dairy cows: a quantitative assessment.

Objectives of this study were to critically review randomized controlled trials, evaluate the effectiveness of supplementation with biotin on milk yield and composition and hoof health in lactating dairy cows, explore sources of heterogeneity among studies, and evaluate publication bias. Quantitative assessments can increase the statistical power with which we study the effect of treatments, such as biotin, on outcomes. A total of 9 papers, with 6 production and 3 hoof health studies, met the eligibility criteria for meta-analysis. Eight studies evaluated various hoof lesions in biotin-supplemented cows that did not meet the inclusion criteria. Eleven comparisons were made of milk production responses to biotin treatment. Data extracted included the number of cows in control and treatment groups, measures of variance of responses (standard error or standard deviation) and P-values. Other data obtained included the duration of treatment before and after calving, parity, breed of cow, type and dose of biotin, delivery method of supplementation, and types of diets. Biotin increased milk production by 1.29 kg per head per day (95% confidence interval=0.35 to 2.18 kg) with no evidence of heterogeneity (I(2)=0.0%). Treatment did not affect milk fat or protein percentages, and a trend to increase fat and protein yields was observed. Milk production and composition results were not influenced by duration of treatment before calving, parity, or diet type. Assessment of biotin supplementation on hoof health indicated that more studies had improved rather than negative or neutral outcomes. The effect of biotin treatment on milk production was relatively large and the effects on fat and protein yields, although not significant, were consistent in direction and magnitude with the milk response. The hoof health responses to biotin should encourage further studies to more effectively define the nature of these responses using consistent criteria for examination of hoof conditions and lameness.

Equine laminitis: induced by 48 h hyperinsulinaemia in Standardbred horses.

REASONS FOR PERFORMING STUDY: Hyperinsulinaemia is known to induce laminitis experimentally in healthy ponies with no history of the condition. Horses are more insulin sensitive than ponies and whether prolonged hyperinsulinaemia and euglycaemia would have a similar laminitogenic effect requires study. OBJECTIVES: To determine if laminitis results when the prolonged euglycaemic hyperinsulinaemic clamp technique (p-EHC) is applied to clinically normal Standardbred horses, and to monitor hoof wall temperature seeking an association between vascular activity and laminitis development. METHODS: Eight young, clinically normal Standardbred horses were assigned into 4 pairs and within each pair, one was assigned randomly to either treatment (n = 4) or control (n = 4) groups. Treated horses received continuous infusions of insulin and glucose until clinical signs of laminitis developed, at which point the horses were subjected to euthanasia. Control horses received an equivalent volume of a balanced electrolyte infusion for the same period. Hoof wall surface temperature (HWST) was monitored continuously throughout the experimental period. RESULTS: All horses in the treatment group were calculated to have normal insulin sensitivity. All treated horses, and none in the control group, developed laminitis (P = 0.01). Pronounced digital pulses were a feature of the treatment group, while insignificant digital pulses occurred in control horses. HWST was higher and less variable in treated horses once hyperinsulinaemia was established. CONCLUSIONS: Healthy Standardbred horses subjected to prolonged hyperinsulinaemia develop laminitis within 48 h, demonstrating that laminitis in horses can be triggered by insulin. POTENTIAL RELEVANCE: Insulin resistance and the associated hyperinsulinaemia place horses and ponies at risk of developing laminitis. This study demonstrates a need for prompt management of the persistent hyperinsulinaemia seen in some endocrinopathies.

Effect of biotin supplementation on claw horn growth in young, clinically healthy cattle.

The effects of orally administered biotin supplementation on the growth of claw horn in young, clinically healthy cattle were analyzed. Twelve, 1-year-old Girolando cattle were randomly assigned to receive either 12.5 mg of diluted powdered biotin (GI) or a control treatment (GII) for 40 consecutive days. Cattle in the GI group showed an average hoof growth of 11.3 +/- 0.72 mm, while those in GII had an average hoof growth of 7.2 +/- 0.78 mm. The results confirmed the positive effect of biotin supplementation on the growth of angle and length of the dorsal hoof wall, hoof sole length, and on resistance to wearing, in young cattle extensively managed.

Efficacy of non-antibiotic treatment options for digital dermatitis on an organic dairy farm.

The objective of this study was to evaluate the efficacy of two non-antibiotic treatment options for digital dermatitis (DD) on an organic certified dairy farm. A randomized clinical trial was conducted using 70 multiparous Holstein cows with an early DD lesion at a USDA certified organic dairy farm in Northern Colorado, USA. Cows were enrolled in the study based on the presence of early DD lesions (scores M1 and M2) and randomly assigned to one of three treatments: (1) topical application of copper sulfate and iodine (CUI); (2) topical application of honey and iodine (HOI); and (3) control subject to no treatment (CON). Cows were evaluated at enrolment and on days 3, 12, 28, and 120 post treatment for pain and lesion size and received a locomotion and a lesion score. Cure was defined as the transition from active to non-active stages (M1/M2 to M0 or M4). The formulations had variable effects on the treatment of DD. The cure rate was numerically higher for CUI on all follow up days. The proportion of cows experiencing pain on d3 after treatment was greater in CON, followed by HOI and CUI. However, this proportion increased in HOI during the follow up period. The CUI group had a greater reduction in lesion size and larger lesions persisted in HOI. Non-antibiotic treatment formulations were partially effective in the treatment of DD in organic dairy cows: The two non-antibiotic formulations resulted in an earlier transition to mature lesions compared with the control group. The CUI combination was the most effective treatment in reducing lesion size, pain, and lameness in affected cows. However, this combination had short-term efficacy, which did not persist throughout the duration of the study. The HOI combination produced only transient reduction in lesion size.

Ex vivo nail infection as an effective preclinical method for screening of new topical antifungals.

Onychomycosis are fungal nail infections comprising of about 50% of onychopathies and are commonly caused by dermatophytes. The treatment of this dermatomycosis requires a long period of time and is associated with high rates of recurrence. In view of the need to evaluate the antifungal performance of promising preclinical compounds, we developed, in this study, a practical and accessibleex vivo model for establishing a Trichophyton rubrum onychomycosis framework using porcine hooves. This model has as its main advantage the similar structural and three-dimensional characteristics that the porcine hooves have with the human nail. The proposed model allowed to evaluate the antifungal activity of a new antifungal compound and a reference drug (terbinafine), both already incorporated into a nail lacquer for topical use. Treatments with compound 3-selenocyanate-indole (Se4a) and with terbinafine incorporated into this nail lacquer completely inhibited fungal growth, corresponding to the profile of in vitro activity observed against T. rubrum. This study concludes that the ex vivo porcine hoof model is an effective alternative method for preclinical screening of drugs or new topical compounds developed to combat onychomycosis. Further studies are needed to compare the permeability of porcine hooves with human nails permeability.

Peripheral blood leukocyte subpopulation of dairy cows with digital dermatitis and effect of hoof trimming with antibiotic treatment.

In the present study, 30 cows were used to evaluate the changes in the peripheral blood leukocyte subpopulation of dairy cows with digital dermatitis (DD) following hoof trimming and antibiotic treatment. The cows were divided into two groups; 18 cows (DD group) had DD on both hind feet, and 12 cows (control group) had four feet with no clinical abnormalities. The DD group was further divided into two groups based on the treatment; the antibiotic group (8 cows) was treated with only 2% lincomycin liquid spray once daily for 3 days, and the trimmed group (10 cows) received trimming of hooves as well as treatment with 2% lincomycin liquid spray. The plasma cortisol concentration was significantly higher in both DD groups before treatment than in the control group, and it decreased significantly after hoof trimming in the trimmed group. The number of CD3(+), CD4(+), WC1(+) and CD21(+) cells in both DD groups before treatment was significantly lower than that of the control group. The number of CD3(+), CD4(+), WC1(+) and CD21(+) cells in the trimmed group increased after treatment. These results indicated that cows with DD suffer from stress and reduced number of T and B cells. Treatment of DD with both hoof trimming and 2% lincomycin liquid spray was effective for reducing the stress and bringing the immune cell number back to the normal range.

Effects of low-dose oligofructose treatment administered via nasogastric intubation on induction of laminitis and associated alterations in glucose and insulin dynamics in horses.

OBJECTIVE: To ascertain whether laminitis can be induced via administration of oligofructose (OF) at doses of 5.0 and 7.5 g/kg in horses and to assess glucose and insulin dynamics before and after treatment. ANIMALS: 19 adult horses. PROCEDURES: Horses were fed OF (1.0 g/kg) mixed with oats for 6 days. Oligofructose at doses of 5.0 and 7.5 g/kg was then mixed with 4 L of water and administered (0 hours) to 8 (group A) and 4 (group B) horses, respectively, via nasogastric intubation; 8 horses received water alone. One horse in group A that did not develop laminitis was subsequently treated again and included in group B. Before and at intervals after treatment, resting plasma glucose and serum insulin concentrations were measured and frequently sampled IV glucose tolerance tests were performed. Area under the glucose curve (AUCg) and area under the insulin curve (AUCi) were calculated, and minimal model analyses were performed. RESULTS: 3 of 8 horses in group A and all 4 horses in group B developed laminitis. Significant treatment-time effects were detected for resting plasma glucose concentrations and AUCg. Among horses in group A, mean AUCg values at 24 and 48 hours were 34% and 32% higher, respectively, than the mean value at 24 hours. Treatment groups did not differ significantly with respect to resting serum insulin concentration, AUCi, or minimal model analysis results. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, laminitis can be induced and glucose dynamics altered via nasogastric administration of 5.0 g of OF/kg. An alteration in insulin dynamics was not detected following treatment with OF.