Effects of in ovo injection of organic selenium on the hatchability of broiler breeder hen eggs and resulting chick physiology and performance.

BACKGROUND: Selenium is an essential mineral for poultry. The conflicting reports about its in ovo injection are the justification for the more detailed investigation. OBJECTIVES: The aim of this study was to investigate the effects of in ovo injection of organic selenium on the hatching traits of broiler chickens and their performance. METHODS: Three hundred and twenty eggs of Ross 308 strain with an average weight of 65 g and 160 chicks were randomly divided into 4 treatment groups (each with 8 replicates of 10 eggs each for hatching parameters and 4 replicates of 10 chicks for broiler farming parameters): negative control (no injection), positive control (in ovo injection of 0.272 mL of normal saline solution) and 2 selenium treatments (in ovo injection of 2.72 or 5.44 µg of organic selenium). Injection was into the amniotic sac on the 10th day of incubation. Effects of in ovo injection on hatching and performance traits, blood parameters, immune responses, carcass characteristics, meat fatty acid profile, cecal microbial population and selenium consternation in the tibia were measured. RESULTS: Fewer chicks from the injected treatments hatched than from the negative control group (p < 0.01). However, the injection of selenium increased feed intake and the final weight of the birds (p < 0.01). Blood parameters were also affected. Glucose and cholesterol in experimental treatment chicks was lower than those of the controls (p < 0.01), whereas blood lipoproteins (VLDL, LDL and HDL) and the ratio of cholesterol to HDL was significantly increased in the treatments injected with selenium (p < 0.01). There was no significant difference in the immune response or microbial population between the experimental groups, but carcass components, such as thigh, breast, wing and abdominal fat weight, were significantly greater in the selenium treatments. CONCLUSIONS: Intra-egg injection of organic selenium produced favourable effects on performance of broiler chickens, although it had no effect on immune response or microbial population. However, the negative effect on hatching of chickens needs to be prevented to result in an acceptable economic return for the producer.

Investigation of the effects of in ovo taurine injection on hatching characteristics and stress reduction potential.

BACKGROUND: In ovo application is the process of administering some nutrients or components into the egg. The main purpose of this application is to ensure that some nutrients are provided to chicks with a short incubation period. Few studies were conducted with taurine in fertile eggs; especially, no observation of hatchability and chick quality has been found. In addition, taurine has an anti-stress impact that fights oxidative factors. OBJECTIVE: To assess the hatchability and chick quality after in ovo taurine administration. To determine the stress that may occur as a result of in ovo application and whether taurine has a stress-reducing effect. METHODS: A total of 1200 fertile eggs from a 34-week-old broiler breeder (Ross 308) flock were categorized into 4 groups with 75 eggs per replicate: control (uninjected), taurine group (0.30 mL dissolved taurine in distilled water), sham control (sterile distilled water) and perforation (eggs perforated and then waxed). On day 14 of incubation, an in ovo injection was administered to the albumen. Data on hatching parameters and hepatic HSP70 levels were obtained using relevant formulas and western blotting, respectively. RESULTS: Control chicks exhibited higher hatchability than other groups, with the taurine group showing the lowest hatchability. The HSP70 levels were the highest in the perforation group compared to the control group. An increase of 21.37% in the taurine group and 83.45% in the sham control group was observed compared to the control group. CONCLUSIONS: The findings suggest that in ovo application may induce increased stress, whereas taurine may have positive effects in mitigating the stress caused by in ovo application.

Carvacrol in ovo delivery optimization and flow dynamics in broiler chicken eggs.

In ovo delivery of carvacrol, the primary active compound in oregano essential oil (OEO) has the potential to enhance gut development in broilers. This study aimed to optimize in ovo application of OEO by investigating day and site of injection and delivery of carvacrol to different embryonic tissues. In Experiment 1, 2 d of injection (embryonic day (E) 12 or 17.5) and 3 sites of injection for OEO (air cell, amniotic fluid, or yolk) were evaluated based on hatchability and posthatching performance. Experiment 2 aimed to examine the impact of combining OEO with the nonionic surfactant polysorbate 80 (p80) at ratios to carvacrol of 0:0, 0:1, 0.5:1, and 1:1 on carvacrol concentration in amniotic fluid, blood, and yolk. The concentration of carvacrol was measured at 3, 6, and 9 h after OEO injection either without (0:1) or with (1:1) p80. Injection of OEO on E12 led to a significant lower hatchability compared to E17.5 (P ≤ 0.01; Δ = 9.2%). Injecting OEO into the air cell, amniotic fluid, or yolk at E17.5 did not significantly affect hatchability and posthatching performance. The highest concentrations of carvacrol found in egg tissues were observed when injected together with surfactant at the 1:1 ratio (P ≤ 0.001; 14.45 µM, 16.64 µM, and 124.82 µM, for air cell, amniotic fluid, and yolk, respectively) compared to the 0:0, 0:1 or 0.5:1 ratios. Carvacrol was highest in the amniotic fluid and blood at the first time point (3 h postinjection) and decreased afterward (P ≤ 0.001), whereas the concentration in yolk remained elevated up to 9 h postinjection. In conclusion, the optimization of the in ovo delivery of carvacrol resulted in that early injection (E12) had negative effects on hatchability and should be avoided. The findings also suggest that using a nonionic surfactant was crucial for an effective delivery of carvacrol in ovo and the migration from amniotic fluid to yolk within 3 h. In addition, carvacrol's persistence in yolk may serve as a route for delivery into the gastrointestinal tract via the yolk stalk during the peri-hatching phase, potentially influencing gut development.

Investigation of a maxillary nerve block technique in guinea pigs (Cavia porcellus): A cadaveric study comparing two injectate volumes.

OBJECTIVE: To investigate and describe an extraoral approach to perform a maxillary nerve block in guinea pigs. STUDY DESIGN: Prospective, randomized, blinded, descriptive, cadaveric study. ANIMALS: A total of 14 adult guinea pig cadavers. METHODS: Two cadavers were used for anatomic dissection and determination of maxillary nerve block approach. A maxillary nerve block via infraorbital approach was then performed in 12 cadavers. A low volume (0.1 mL) or high volume (0.2 mL) of diluted methylene blue injectate was randomly assigned to the right or left side, with the other volume used for the contralateral side. The maxillary nerve was dissected after each injection by an investigator blinded to injectate volume. The region of dye distribution was identified, and the degree of staining assigned an accuracy score (0-2). Nerve coverage was considered adequate if ≥6 mm of circumferential staining was present. RESULTS: Accuracy evaluation indicated successful dye deposition in 10/12 [2 (0-2), median (range)] injections in the low volume group and 8/12 [2 (1-2)] injections in the high volume group. The majority (79.2%) of injections resulted in adequate nerve staining. There were no statistically significant differences between injectate volumes for accuracy (p = 0.64) or adequacy (p > 0.99) of staining. CONCLUSIONS: The infraorbital approach is a simple and practical method for maxillary nerve blockade in guinea pigs. An injectate volume of 0.1 mL results in adequate maxillary nerve coverage; however, additional studies are needed to assess the efficacy in clinical use.

Accuracy of drawing up liquid medications by veterinary anaesthetists and nurses.

OBJECTIVE: To evaluate the accuracy with which veterinary anaesthetists and nurses of different experience draw up small volumes of liquid. STUDY DESIGN: Prospective blinded randomized study. METHODS: A group of seven European College of Veterinary Anaesthesia and Analgesia (ECVAA) diplomates, six ECVAA residents and six anaesthesia nurses participated in the study. For each participant, five target volumes between 0.01 and 1.0 mL were randomly allocated. They were drawn up 20 times in random order using a 1 mL syringe attached to a 23 gauge needle. A total of 1900 measurements were analysed. An investigator filled the syringe and needle dead space with water for injection before each experiment. The change in mass of the syringe once filled to the target volume was used to calculate the actual volume of liquid drawn up. RESULTS: Large errors were made with measurements < 0.04 mL, so they were excluded from the statistical analysis. There was a significant effect of the target volume (p < 0.001) and of the order (p < 0.01). For each mL increase in the target volume, the absolute error decreased by 4% and from the first to the hundredth withdrawal each participant's absolute error decreased by 1%. The target volume was underestimated in 52.3% of measurements, 42% were overestimated and 5.7% were on target. None of the between-subject predictor variables approached significance. CONCLUSIONS AND CLINICAL RELEVANCE: Accuracy of veterinary anaesthetists and nurses in drawing up medications decreases as the target volume becomes smaller. Small veterinary patients receive small volumes of anaesthetic drugs with higher risks of overdosing compared with larger dogs. Years of experience and staff grade are not associated with greater accuracy. Large percentage errors may be seen with target volumes less than 0.04 mL. Dilutions are recommended for volumes > 0.19 mL in 1 mL syringe.

Decreasing polymethyl methacrylate bone cement concentration extends working and setting times in vitro.

OBJECTIVE: To determine setting and temperature properties of diluted polymethyl methacrylate (PMMA) bone cement in vitro to assess utility for vocal fold augmentation in horses. SAMPLES: 4 dilutions of PMMA equivalent to volumes of 15 mL, 20 mL, 25 mL, and 30 mL PMMA powder (PMMAp) in 10 mL solvent. METHODS: For each volume PMMAp, setting times (tset), peak temperatures (Tmax), and times to peak temperature (tmax) were determined using a temperature data logger in a 4-mL volume of PMMA. Injectability was assessed in vitro by documenting the force required to inject 0.2 mL PMMA through an 18-gauge 3.5-inch spinal needle attached to a 6-mL syringe at 1-minute intervals. Working time (twork) was calculated from a linear regression of injectability. RESULTS: Peak temperatures increased with increasing volume of PMMAp: 56 °C, 86 °C, 99 °C, and 101 °C. Times for tset, twork, and tmax were inversely proportional to PMMA concentrations, resulting in tset of 23, 21, 17, and 14 minutes; twork of 22.75, 12.25, 7, and 4 minutes; and tmax of 28, 24, 19, and 16 minutes, respectively, for 15, 20, 25, and 30 mL PMMAp. Pairwise comparisons for all analyses were significant apart from Tmax for 25 and 30 mL PMMAp (P = .96) and twork for 20 and 25 mL PMMAp (P = .06). CLINICAL RELEVANCE: Decreasing the concentration of PMMA bone cement resulted in longer working times and setting times; however, peak temperatures did not differ between the 2 strongest concentrations. Further research is warranted to quantify diluted PMMA properties for in vivo use for vocal fold augmentation in horses.

In-Ovo injection of melittin into Alexandria chicken eggs: a way for early immune acceleration.

This study intended to assess the properties of in-ovo administration of Melittin (MLT) on hatchability, chick yield, hematology, immunological indices and relative organs weight of Alexandria chickens at hatch. A total of 600 eggs with an average weight of (45.12 g), were gathered and split into five groups: a non-injected group or negative control (NC), a saline injection group or positive control (PC), and three concentrations of MLT (5, 10 and 15 µg of MLT per egg, respectively). On day 18 of incubation, eggs from the injection groups were injected into the amniotic fluid from the large end with the in-ovo injection solutions (0.2 ml per egg). Results indicated that 10 µg MLT/egg positively affected the weight and yield of chicks. In addition, our findings indicated that the in-ovo administration with 10 or 15 µg MLT/egg was superior in most of the immunological indicators (spleen and bursa relative weights, immunoglobulins IgG and IgM, T cells and B cells). In conclusion, in order to improve the immune efficiency (early immune acceleration) of Alexandria chicks, which may contribute to offering a significant boost to their future performance, this study suggests injecting eggs with 5 or 10 µg MLT/egg.

The investigation of the effects of vitamin A, vitamin E, and ß-carotene plus vitamin E on some fertility parameters in ewes.

This study was aimed at investigating the effects of vitamin A (VITA), vitamin E (VITE), and combined ß-carotene plus vitamin E (ßCAR+VITE) injections on some fertility parameters in ewes. Estrus synchronization was performed by treating the ewes with intravaginal FGA sponges impregnated with 30 mg of fluorogestone acetate. On the days of the insertion and withdrawal of the intravaginal sponges, groups VITA, VITE, and ßCAR+VITE were administered with 500 000 IU of vitamin A, 50 mg of vitamin E, and a combination of ß-carotene plus vitamin E, respectively. The ewes in the control group (C) were maintained for control purposes. Statistically significant differences were determined between groups VITA and ßCAR+VITE, groups VITE and ßCAR+VITE, and groups C and ßCAR+VITE, as well as groups VITE and C, groups VITA and C for the multiple birth rates. While significant differences were determined between groups VITA and C, groups VITE and C, and groups ßCAR+VITE and C for the lambing rates, it was ascertained that the ratio of newborn lambs to delivered ewes (litter size) significantly differed between groups VITA and ßCAR+VITE, groups VITA and C, groups VITE and ßCAR+VITE, groups VITE and C, and groups ßCAR+VITE and C. The highest MDA level and lowest GSH level were determined on day 20 after mating in the control group. In conclusion, it is suggested that both multiple birth rates and litter size can be increased by the combined administration of ß-carotene and vitamin E.

Blind Versus Ultrasound-Guided Low-Volume Perineural Injection of Tibial and Fibular Nerves in Equine Cadaver Limbs.

Techniques for local anesthesia of the tibial (TN) and superficial and deep fibular nerves (FNs) in horses are well established. Ultrasound-guided perineural blocks can identify the nerve location, reduce the anesthetic volume needed and avoid needle misplacement. The aim of this research was to compare the success of blind perineural injection technique (BLIND) to ultrasound-guided technique (USG). Fifteen equine cadaver hindlimbs were divided into two groups. Perineural injection of the TN and FNs was performed using a mixed solution of radiopaque contrast, saline and food dye. BLIND (n = 8) used 15 mL for the TN and 10 mL for each fibular nerve. USG (n = 7) used 3 mL for the TN and 1.5 mL for each fibular nerve. The limbs were radiographed immediately after injections and sectioned transversally to evaluate the diffusion and presence of the injectate adjacent to the TN and FNs. The presence of dye immediately adjacent to the nerves was considered a successful perineural injection. No statistically significant difference was observed between groups for success. Distal diffusion of injectate following perineural injection of the TN was significantly less for USG compared to BLIND. Proximal, distal and medial diffusion of injectate following perineural injection of FNs was significantly less for USG compared to BLIND. Low-volume USG results in less diffusion but similar success compared to BLIND leaving it up to veterinarian preference when selecting a technique.

Evaluation of Survival of Subdermal Plexus Skin Flaps in 8 Cats After Injection of Platelet Rich Plasma.

The aim of the present study was to evaluate the effect of platelet-rich plasma injection on the survival of experimentally developed subdermal plexus skin flaps in cats. Two flaps, 2 cm wide and 6 cm long were created bilaterally in the dorsal midline in 8 cats. Each flap was randomized to 1 of 2 groups: platelet-rich plasma injection and control. After flap development, the flaps were immediately placed back on the recipient bed. Then 1.8 mL of platelet-rich plasma was equally injected into 6 different parts of the treatment flap. All flaps were evaluated macroscopically daily and on days 0, 7, 14, and 25 by planimetry, Laser Doppler flowmetry, and histology. Mean flap survival on day 14 was 80.437 % (±22.745) for the treatment group and 66.516 % (±24,12) for the control group with no statistically significant difference between the 2 groups (P = .158). Histologically, a significant difference was found between the base of the PRP and the control flap in edema score on day 25 (P = .034). In conclusion, there is no evidence to support the use of platelet-rich plasma in subdermal plexus flaps in cats. However, the use of platelet-rich plasma may aid in reducing edema of subdermal plexus flaps.

Efficacy of in ovo delivered resveratrol (Trans 3, 4, 5-trihydroxystilbene) on growth, carcass weights, and blood metabolites of broiler chicks.

This research aimed to determine the impact of in ovo resveratrol (RV) (Trans.3, 4, 5-trihydroxystilbene) injection on performance, carcass weights, and blood constituents in Cobb chicks. A total of 300 fertile broiler breeder eggs were divided into five treatments as 60 eggs in each group, with 6 replicates (10 eggs each). The groups were as follows: (1) negative control (without additive), (2) positive control (saline injection), (3) 1.5 mg RV, 4) 3.0 mg RV, and (5) 4.5 mg in ovo RV injections. Significant changes were found in the amount of forage consumed per day, and the feed conversion ratio between treatment groups. RV injection significantly altered the parameters of the complete lipid profile, including total cholesterol, triglycerides, low-density lipoproteins, and very low-density lipoproteins, in plasma. RV injection significantly affected blood biochemical parameters, including total protein, albumin, and globulin in broiler chicken plasma. Liver and kidney functions were influenced by the injection of RV eggs into broiler chickens. The antioxidant enzymes also changed significantly after RV injection. Moreover, in egg injection from RV the immunogenicity (IgG and IgM) in broiler chickens were significantly altered. In conclusion, in ovo injection with RV significantly improved liver and kidney functions, antioxidant activity, and immune function in Cobb broilers.

Determination of the embryotoxic effect of maropitant using an in ovo model.

The aim of this research was to determine the embryotoxic and teratogenic effects and lethal dose (LD50) of maropitant in ovo, using fertile chicken eggs. The study was designed in two stages, CHEST-I and CHEST-II. For CHEST-I, 210 fertile eggs were divided into seven equal groups; control, saline solution and 5 different doses of maropitant (10, 5, 2.5, 1.25, 0.625 mg/kg) injected groups. For CHEST-II, 150 fertile eggs were divided into five equal groups; control, saline solution and 3 different doses of maropitant (8, 6, 4 mg/kg)-injected groups. Eggs were opened on day 21 of incubation. Maropitant did not cause teratogenicity at any dose, while higher embryonic death rates were observed at doses above 4 mg/kg. The LD50 dose of maropitant was determined as 7.24 mg/kg. In conclusion, maropitant should only be used after full consideration of risks and benefits in pregnancy.

A case of priapism following intrathecal morphine injection in a dog.

Background: Priapism refers to prolonged erection unrelated to sexual stimulation, with severe sequelae unless treated. In humans, it is a rare complication associated with epidural or spinal opioid administration. Its pathophysiology is unclear. This is the first report of priapism following neuraxial anesthesia in dog. Case Description: An intrathecal morphine injection (30 mcg/kg) at L5-L6 for postoperative analgesia was given at the end of surgery for removal of cutaneous mastocytomas of the abdomen and left axillary lymphadenectomy. Painless penile erection occurred 2 hours later and lasted 6 hours, before spontaneously resolving 7-8 hours after the injection. No pain or other adverse events (e.g., nausea, urinary retention, and itching) were recorded. Recovery was complete without treatment. Conclusion: Painless, self-resolving priapism is a rare complication associated with intrathecal morphine injection in dogs.

Trichloroacetic Acid Injection for Treatment of Eyelid Apocrine Hidrocystomas in a Cat.

Apocrine hidrocystoma (AH) is a benign cystic lesion infrequently reported in the eyelids of cats. There are several reports of application of trichloroacetic acid (TCA) for treatment of eyelid apocrine hidrocystomas with high success rates in humans. This is the first report of intralesional injection of TCA for the treatment of eyelid AH in a feline. A 12-year-old Persian female spayed cat has been presented for evaluation of cystic masses on the eyelids. The cat had signs of ocular discomfort and two large cysts on the lower eyelid of the right eye. Ophthalmic examination was suggestive of eyelid AH. The cat underwent general anesthesia, and TCA 20 % was injected intracystically. The diagnosis of AH was confirmed by cytology. Two years later, there was no recurrence of the cyst. This case highlights the successful treatment of a large AH in the eyelids of a cat with TCA. Findings suggest that chemical ablation with TCA may be a useful treatment for AH in cats.

Economic impact of gastrointestinal nematodes in Morada Nova sheep in Brazil / Impacto econômico de nematódeos gastrintestinais em ovinos Morada Nova no Brasil

Abstract This study evaluated the economic impact of gastrointestinal nematode (GIN) infection in Morada Nova lambs under different parasite chemical control conditions. For this, 246 lambs, in the rainy and dry season, were randomized into groups according to their anthelmintic treatment with levamisole: control (CT: no treatment); routine treatment (RT: treated every 42 days); and targeted selective treatment (TST: treated according to the average daily weight gain, DWG). From 63 days of age (D63) to D210, the lambs were weighed and monitored for GIN infection parameters. Spending on anthelmintics in the production system was 1.3% of the total economic result. The economic result per animal (R$ 5.00 = US$ 1.00) was higher in the RT group, amounting to US$ 6.60 in the rainy and US$ 5.69 in the dry season, due to higher DWG. Thus, RT presented economic results 14.4% and 10.9% higher than CT, and 7.2% and 1.9% higher than TST, in the rainy and dry season, respectively. However, fast development of resistance made RT unfeasible. Here, the economic impact of GIN infection on a national scale is discussed, demonstrating its importance and the impossibility of profitable and sustainable sheep production without adequate control.

Resumo Este estudo avaliou o impacto econômico da infecção por nematoides gastrintestinais (NGI), em cordeiros Morada Nova, sob diferentes condições de controle químico dos parasitas. Para isso, 246 cordeiros, na estação chuvosa e seca, foram randomizados em grupos de acordo com o tratamento com levamisol: controle (TC: sem tratamento); tratamento rotineiro (TR: tratado a cada 42 dias); e tratamento seletivo direcionado (TST: tratado de acordo com o ganho de peso médio diário, GMD). Dos 63 dias de idade (D63) ao D210, os cordeiros foram pesados ​​e monitorados quanto aos parâmetros de infecção por NGI. O gasto com anti-helmínticos no sistema produtivo foi de 1,3% do resultado econômico total. O resultado econômico por animal (R$ 5,00 = US$ 1,00) foi maior no grupo RT, totalizando US$ 6,60 na estação chuvosa e US$ 5,69 na seca, devido ao maior GMD. Assim, o RT apresentou resultados econômicos 14,4% e 10,9% superiores ao TC, e 7,2% e 1,9% superiores ao TST, no período chuvoso e seco, respectivamente. Entretanto o rápido desenvolvimento de resistência inviabiliza o TR. O impacto econômico da infecção por NGI em escala nacional são aqui discutidos, demonstrando sua importância e a impossibilidade de uma ovinocultura lucrativa e sustentável sem o controle adequado.

The use of intrarenal injection of sodium pentobarbital for euthanasia in cats.

OBJECTIVE: To determine whether intrarenal injection of sodium pentobarbital is a viable method for euthanasia in anesthetized client-owned cats and assess potential factors associated with time to cardiopulmonary arrest (TCPA) for such treated cats. ANIMALS: 131 client-owned cats. PROCEDURES: In this retrospective study, client-owned cats presented for euthanasia between March 1, 2009, and January 15, 2010, were evaluated by veterinarians to determine suitability of intrarenal injection versus other methods of euthanasia. Cats included were anesthetized and then received 6 mL of sodium pentobarbital (390 mg/mL) by intrarenal injection. Results for TCPA were compared for cats grouped on the basis of variables of interest. RESULTS: 131 cats were included, of which 74 (79%) had a TCPA < 1 minute and 28 (21%) had a TCPA between 1.5 and 8 minutes after intrarenal injection. Most (124/131 [95%]) cats had no observable reaction to the intrarenal injection other than cardiopulmonary arrest. Median TCPA was longer for cats without (1 min; 25/131 [19%]) versus with (0 min; 106/131 [81%]) palpable kidney swelling upon injection. CLINICAL RELEVANCE: The effects of intrarenal injection of sodium pentobarbital in cats of the present study were similar to those typically observed with IV administration of euthanasia solution. When this intrarenal injection method is used, cardiopulmonary arrest with few agonal reactions can be expected to occur quickly in most patients. The intrarenal injection method is suited for euthanasia of anesthetized cats with easily located kidneys when IV access may be difficult.

Transdermal Application of Anesthetic Preparations Is Effective in Increasing Mechanical Nociceptive Threshold at Perineural Injection Sites in Horses.

Perineural anaesthesia is an essential part of lameness investigation in horses. Painful reactions to injections imply a high risk of accidents for veterinarians. To evaluate the efficacy of two topical anaesthetic creams on desensitization of the skin at two perineural injection sites and to evaluate the effects these formulations may have on pain perception distally to the application sites. Randomized, blind, prospective study. 14 healthy horses were randomly divided in two groups of 7 horses each, one treated with EMLA and one with PLIAGLIS preparations. The mechanical nociceptive threshold (MNT) was recorded via pressure algometer at the baseline and one hour after application (T1) on palmar and palmar digital nerve blocks sites. Skin sensation was also assessed on the distal limb, recording MNT on the coronary band and on both bulbs of the heels. Data were compared with a 3-way ANOVA statistical analyses. No significant difference was encountered in the MNT values between baseline records and records at T1 at the control sites. A significant difference of MNT values was detected between the two time points in the treated limbs. The significant sources of variation in our study were the time and the presence of anaesthetic cream. The type of treatment did not determine any differences. Furthermore, no significant difference of MNT values at sites distal to treatment was recorded between the two time points, both in the treated limb and control limbs. Both formulations are effective to obtain superficial skin desensitization.