Reduction in cytoplasmic lipid content in bovine embryos cultured in vitro with linoleic acid in semi-defined medium is correlated with increases in cryotolerance.

UNLABELLED: We examined whether culturing embryos with linoleic acid (LA) in semi-defined medium reduces lipid accumulation and improves cryosurvival after vitrification. Embryos were cultured with LA (100 µM) and a semi-defined medium was used during in vitro culture (IVC), in which the fetal calf serum was substituted by bovine serum albumin (BSA). There was a reduction (P < 0.05) in the embryonic development rate ( CONTROL: 25.8% versus LA: 18.5%), but the proposed system was effective in promoting the decrease (P = 0.0130) in the intracellular lipid content ( CONTROL: 27.3 ± 0.7 versus LA: 24.6 ± 0.7 arbitrary fluorescence units of embryos stained with the fluorescent dye Nile Red), consequently increasing (P = 0.0490) the embryo survival after 24h of culture post-warming ( CONTROL: 50.0% versus LA: 71.7%). The results question the criteria used to evaluate the efficiency of an in vitro production system specifically with relation to the maximum number of blastocysts produced and suggest that might be more appropriate to improve the desired characteristics of embryos generated in accordance with the specific purpose of in vitro embryo production, commercial or scientific. In conclusion, supplying LA to serum-free culture medium was found to adversely affect the rates of embryo development to the blastocyst stage, but significantly reduced embryo lipid accumulation and improved cryopreservation survival.

Seroepidemiology of Toxoplasma gondii, Neospora caninum, and Leishmania spp. infections and risk factors for cats from Brazil.

The seroprevalence of infection by Toxoplasma gondii, Neospora caninum, and Leishmania spp. was detected through an indirect immunofluorescence in 70 cats from the Andradina Municipality, São Paulo State, Brazil. Anti-T. gondii antibodies (titer >64) were detected in 15.7% (11/70) of animals, whereas positivity for N. caninum (titer 16) was not observed in any animal. Of the cats from urban and rural areas, 10.4% (5/48) and 27.2% (6/22) were positive for T. gondii, respectively. Breed, age, food, and contact with animals of other species were significant for considering the positivity for T. gondii (P ≤ 0.0001). Cats having access to streets (17.1%, 11/64), cats cohabiting with rats (19.6%, 10/51), and cats feeding on homemade food and raw milk (27.2%, 6/22) were positive for T. gondii. In addition, 4.2% (3/70) of the cats were positive for Leishmania spp. by ELISA technique and negative by IFAT without coinfection with T. gondii and Leishmania spp. There was no serological positivity against feline immunodeficiency virus or feline leukemia virus. In conclusion, T. gondii infection in part of the feline population from Andradina is not linked to immunosuppressions or coinfections but probably to postnatal infection in association with the type of diet and presence of rats.

Effect of intravenous propofol and remifentanil on heart rate, blood pressure and nociceptive response in acepromazine premedicated dogs.

OBJECTIVE: To investigate the cardiorespiratory, nociceptive and endocrine effects of the combination of propofol and remifentanil, in dogs sedated with acepromazine. STUDY DESIGN: Prospective randomized, blinded, cross-over experimental trial. ANIMALS: Twelve healthy adult female cross-breed dogs, mean weight 18.4 ± 2.3 kg. METHODS: Dogs were sedated with intravenous (IV) acepromazine (0.05 mg kg(-1) ) followed by induction of anesthesia with IV propofol (5 mg kg(-1) ). Anesthesia was maintained with IV propofol (0.2 mg kg(-1) minute(-1) ) and remifentanil, infused as follows: R1, 0.125 µg kg(-1) minute(-1) ; R2, 0.25 µg kg(-1) minute(-1) ; and R3, 0.5 µg kg(-1) minute(-1) . The same dogs were administered each dose of remifentanil at 1-week intervals. Heart rate (HR), mean arterial pressure (MAP), respiratory rate (f(R) ), end tidal CO(2) (Pe'CO(2) ), arterial hemoglobin O(2) saturation, blood gases, and rectal temperature were measured before induction, and 5, 15, 30, 45, 60, 75, 90, and 120 minutes after beginning the infusion. Nociceptive response was investigated by electrical stimulus (50 V, 5 Hz and 10 ms). Blood samples were collected for plasma cortisol measurements. Statistical analysis was performed by anova (p<0.05). RESULTS: In all treatments, HR decreased during anesthesia with increasing doses of remifentanil, and increased significantly immediately after the end of infusion. MAP remained stable during anesthesia (72-98 mmHg). Antinociception was proportional to the remifentanil infusion dose, and was considered satisfactory only with R2 and R3. Plasma cortisol concentration decreased during anesthesia in all treatments. Recovery was smooth and fast in all dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Infusion of 0.25-0.5 µg kg(-1) minute(-1) remifentanil combined with 0.2 mg kg(-1) minute(-1) propofol produced little effect on arterial blood pressure and led to a good recovery. The analgesia produced was sufficient to control the nociceptive response applied by electrical stimulation, suggesting that it may be appropriate for performing surgery.

Dog culling and replacement in an area endemic for visceral leishmaniasis in Brazil.

Measures employed to control visceral leishmaniasis in Brazil have focused on vector control by residual insecticide spraying and diagnosis of infection with elimination of positive dogs. We describe dog culling and replacement in a Brazilian endemic area (the Alvorada District, Araçatuba, SP) in order to better understand dog population dynamics when elimination of the dog reservoir is adopted as the main control measure. From August 2002 to July 2004, 60.9% of the estimated dog population for the area was culled with a mean age of 34 months old. The presence of anti-Leishmania sp. antibodies was recorded for only 26.7% of the euthanized canines. Replacement was observed in 38.8% of the cases, some of them by 2 or more dogs and in a mean time of 4 months. Dogs were replaced mostly by puppies of both sexes with a mean age of 6.8 months. From August 2002 to April 2005 we were able to follow-up 116 of these dogs, during a mean time of 8.7 months. Canine visceral leishmaniasis seropositivity by ELISA was observed in 42.2% of the followed dogs, 30.6% of which were already positive at the first evaluation. By the end of the follow-up period 37% of the dogs were submitted to euthanasia, with a mean age of 18.3 months. In the studied CVL endemic area of Brazil, euthanasia and the subsequent replacement ratio were high, increasing the dog population turnover and leading to a younger population that might be more susceptible to a variety of other infectious diseases in addition to CVL. Dog culling as a control strategy for VL should be reassessed.

Comparative evaluation of enzyme-linked immunosorbent assay based on crude and purified antigen in the diagnosis of canine visceral leishmaniasis in symptomatic and oligosymptomatic dogs.

This study evaluated the performance of crude total antigen (CTA) and fucose-mannose ligand antigen (FML) in an enzyme-linked immunosorbent assay for diagnosis of canine visceral leishmaniasis (CVL). The assays used sera from known negative controls (n=30), clinically symptomatic (n=30) and oligosymptomatic (n=30) parasitologically proven infection (by microscopy). Aspirates of popliteal lymph node from infected canines were colleted to score parasitism and compared with the ELISA results. The study indicated that FML used in ELISA provided high sensitivity for detecting oligosymptomatic dogs (90%) and CTA showed greater sensitivity than FML for symptomatic canines (90%). In oligosymptomatic dogs, specificity was 100% for CTA-ELISA, but in symptomatic dogs, FML specificity was higher (96.7%) than CTA-ELISA (93.3%). A significant correlation was observed between the degree of parasitism and the results obtained in CTA-ELISA. Since no available antigen offers 100% specificity and sensitivity for CVL diagnosis, the choice of antigen used must depend on the aim of the investigation.

[A descriptive profile of the canine population in Araçatuba, São Paulo State, Brazil, from 1994 to 2004]. / Estudo descritivo da estrutura populacional canina da área urbana de Araçatuba, São Paulo, Brasil, no período de 1994 a 2004.

From 1994 to 2004, the canine population in Araçatuba, São Paulo State, Brazil, suffered two major canine zoonoses: rabies and visceral leishmaniasis. Changes in the dog population during this period were evaluated using canine census data from 1994 and 2004 and the results of blood samples for diagnosis of canine visceral leishmaniasis in 1999. The ratio of dogs per 10 inhabitants varied from 1.7 in 1994 to 2.0 in 1999 and 1.8 in 2004. The percentage of puppies less than 1 year of age increased from 20% to 32.5%, and the number of euthanized dogs also increased after 1999, when visceral leishmaniasis began to appear. The number of dogs and percentage of puppies varied between different areas of the city, and neighborhoods with a higher percentage of young animals showed more cases of both human and canine leishmaniasis. This result may be due to control measures applied in these areas in response to cases of human and canine visceral leishmaniasis, but the increase in the younger canine population can be accompanied by increased susceptibility in these animals, thus favoring maintenance of the disease in the area.

Morphological aspects of tympanic bulla after lateral osteotomy in cats.

PURPOSE: To evaluate tympanic bulla healing after experimental lateral osteotomy in cats. METHODS: Twenty adult cats were submitted to unilateral lateral bulla osteotomy and divided into two groups: cats of B1 group (n=10) were euthanized at 8 weeks and cats of B2 group (n=10), at 16 weeks postoperative. RESULTS: Oblique lateral radiographs taken immediately postoperative showed interruption in the contour of the external acoustic meatus of the operated bullae in all cats of both groups (McNemar test: p=0.0010*). This feature was still observed in the radiographs taken after 8 and 16 weeks postoperative (McNemar test: B1 p=0.0020*; B2 p=0.0312*). Macroscopic examination showed that the operated bullae were similar to the normal ones, with preservation of the tympanic cavity. Connective tissue at the osteotomy site was significantly found in the operated bullae in both groups (McNemar test: B1 p=0.0020*; B2 p=0.0010*). The length of connective tissue at the osteotomy site was measured by histomorphometry. There was no statistically significant difference between the values of B1 group and B2 group (Mann-Whitney test: p=0.0524). CONCLUSIONS: Experimental lateral osteotomy did not alter significantly the tympanic bulla conformation and complete regeneration of the tympanic bulla frequently did not occur before 16 weeks of postoperative period.

Does lipid peroxidation and oxidative DNA damage differ in cryopreserved semen samples from young, adult and aged Nellore bulls?

The aims of this study were to evaluate cryopreserved semen of Nellore bulls of different ages and verify whether sperm quality declines with advancing age and whether lipid peroxidation and DNA damage are involved in this process. For this purpose, 40 Nellore bulls were divided into three age groups: Young, aged 1.8-2 years (n = 9); Adult, aged 3.5-7.0 years (n = 19); and Seniors, aged 8.0-14.3 years (n = 12). Three ejaculates were collected from each bull, cryopreserved and evaluated for various parameters including membrane integrity, mitochondrial potential (FITC-PSA and JC1), lipid peroxidation (C-11BODIPY 581 / 591) and oxidative DNA damage (8OHdG) using flow cytometry. The thawed semen of senior bulls was characterized by a low percentage of motile sperm (33.7 ±â€¯6.1%), higher damage to the plasma and acrosomal membrane (37.5 ±â€¯9.8%), and low mitochondrial potential (29.1 ±â€¯13.8%), as well as higher percentages of peroxidated cells (53.6 ±â€¯12.2%) and DNA damage (44.1 ±â€¯11.0%; P < 0.05). Lipid peroxidation was negatively correlated with motility (r = -0.35, P < 0.0002), average mitochondrial potential (r = -0.42; P < 0.0001) and showed a positive correlation with membrane injury and oxidative DNA damage (r = 039; P = 0.0003). Young bulls presented superior thawed sperm quality, possibly due to greater resistance to oxidative stress and, consequently, to cryopreservation. In conclusion, the sperm quality of bull semen declines with advancing age and is strongly associated with increased oxidative damage to both the plasma membrane and DNA.

Factors associated with positivity for canine visceral leishmaniosis in an endemic area in Brazil.

Visceral leishmaniosis (VL) is a public health problem and its occurrence depends primarily on the presence of the vector and susceptible hosts; in the urban environment, the dog is the main reservoir. This study aimed to analyze the distribution of canine visceral leishmaniosis (CVL) and factors associated with it in an urban area endemic for VL. Analysis of the variables was based on 2755 epidemiological records of dogs positive and negative for CVL over a three-year period (2009, 2010 and 2011). A dog was considered positive when it presented amastigotes in the cytological examination of lymph node biopsy and/or was seropositive by immunoenzymatic and indirect immunofluorescence assays. CVL positive dogs were observed throughout the town, but significant differences were observed between the sectors analyzed (P<0.0001), with two sectors showing higher positivity. CVL prevalence was 35.9% and was significantly associated with age and breed (P<0.0001). Concerning symptoms, 44.3% of symptomatic dogs were positive for LV (P<0.0001) in an urban area endemic for this zoonosis.

Clinical and parasitological evaluation of dogs naturally infected by Leishmania (Leishmania) chagasi submitted to treatment with meglumine antimoniate.

Aiming to evaluate the efficacy of the treatment of canine visceral leishmaniasis, to verify the occurrence of a possible disease relapse, and to search for the presence of the parasites after the end of the treatment, seven dogs naturally infected by Leishmania (Leishmania) chagasi were used. The dogs were subjected to a treatment with 75 mg/kg meglumine antimoniate subcutaneously every 12 h for 21 days, and followed-up for a period of 6 months. During the whole experimental period the animals wore deltamethrin collars and were kept in a screened kennel to avoid reinfection. Lymph node and bone marrow aspiration biopsy was carried out to search for the parasite at seven moments: before the treatment, 30, 60, 90, 120, 150 and 180 days after the start of the treatment. After the end of the experiment all dogs were humanely euthanized. Then, spleen and liver "imprints" and in vitro cultures were carried out to search for amastigote forms of the parasite. During the treatment all animals presented remission of symptoms. However, two dogs were observed to present new symptoms in the course of the experiment. At the end of the experiment, the presence of amastigote forms of the parasite was evidenced in five of the seven dogs. This enabled us to conclude that the treatment promoted clinical cure but did not eliminate the parasites completely.

[Dog replacement in an area endemic for visceral leishmaniasis]. / Reposição de cães em área endêmica para leishmaniose visceral.

This study aimed to estimate the dog replacement rate in an area endemic for visceral leishmaniasis, in which slaughter of seropositive animals was indicated as a control measure, and to evaluate the reasons why new animals were or were not acquired. The animals were replaced in 44.5% of the cases, and this was done mainly because of the need for a companion or guard dog. The main reason for not replacing the dog was fear of visceral leishmaniasis.

Age structure of owned dogs under compulsory culling in a visceral leishmaniasis endemic area.

The age structure of the dog population is essential for planning and evaluating control programs for zoonotic diseases. We analyzed data of an owned-dog census in order to characterize, for the first time, the structure of a dog population under compulsory culling in a visceral leishmaniasis endemic area (Panorama, São Paulo State, Brazil) that recorded a dog-culling rate of 28% in the year of the study. Data on 1,329 households and 1,671 owned dogs revealed an owned dog:human ratio of 1:7. The mean age of dogs was estimated at 1.73 years; the age pyramid indicated high birth and mortality rates at the first year of age with an estimated cumulative mortality of 78% at the third year of age and expected life span of 2.75 years. In spite of the high mortality, a growth projection simulation suggested that the population has potential to grow in a logarithmic scale over the years. The estimated parameters can be further applied in models to maximize the impact and minimize financial inputs of visceral leishmaniasis control measures.

Effect of age and abomasal puncture on peritoneal fluid, hematology, and serum biochemical analyses in young calves.

The goals of this study were to evaluate techniques for collection of peritoneal fluid from calves, establish reference ranges for fibrinogen in peritoneal fluid during the 1st month of life, and determine if abomasal puncture would alter peritoneal fluid or hematologic variables. Twenty-two healthy Holstein calves underwent 3 peritoneal fluid collections on day 1, day 15, and day 30 of age. Fibrinogen concentration in peritoneal fluid was 0.20 g/dL and 0.10 g/dL (P < .05) for day 1 and day 30, respectively, and 0.10 at day 15 (P > .05) for calves without abomasal puncture. Plasma fibrinogen concentration was 0.60 g/dL and 0.70 g/ dL (P < .05) for days 15 and 30, respectively, in calves without abomasal puncture. There were no significant differences (P < or = .05) in peritoneal fluid and peripheral blood total protein and fibrinogen concentrations, specific gravity, total and differential cell count, or erythrocyte counts between calves with or without abomasal puncture. We concluded that the reference ranges established for fibrinogen and total protein concentration are important for accurate evaluation of peritoneal fluid in calves for further comparison with similar-aged animals with gastrointestinal-tract or abdominal-cavity disease. Additionally, accidental abomasal puncture does not alter values of fibrinogen, total protein, and nucleated cell count in peritoneal fluid and does not cause apparent clinical abnormalities.

Tissue apoptosis in mice infected with Leptospira interrogans serovar Icterohaemorrhagiae.

BACKGROUND: This investigation aimed to evaluate the occurrence of some apoptotic features induced by Leptospira interrogans serovar Icterohaemorrhagiae infection in young BALB/c mice during 2, 4, 7, 10, 14 and 21 days post-infection (dpi). METHODS: The animals were euthanized and lung, liver and kidneys were harvested to histopathology analysis and immunohistochemistry to caspase-3 antigen detection was performed. RESULTS: Chromatin condensation in kidney and liver tissues, but not in lung tissue, was observed. Caspase-3 reactive cells, mainly characterized as renal epithelial cells, were detected in the days 14 and 21 at high levels when compared to days 2, 4 and 7 (p = 0.025; p < 0.05). Lung sections revealed caspase-3 labeled alveolar cells in 10 and 14 days post-infection was higher than observed at 7 days (p = 0.0497; p < 0.05). Liver sections demonstrated reactive cells at a highest level at 14 and 21 days post-infection when comparison to 2, 4, 7 and 10 days (p = 0.0069; p < 0.05). CONCLUSIONS: Our results suggest that infection of L. interrogans induce in kidney, liver and lung an activation of apoptosis mediated by caspase-3 dependent pathway in later phases of infectious process.

Canine rabies epidemiology in Araçatuba and neighborhood, Northwestern São Paulo State-Brazil.

Epidemiological characteristics of canine rabies in the northwest region of São Paulo State (Araçatuba region), Brazil, from 1993 to 1997 are presented. Out of 1,984 dogs, a total of 351 (17.7%) were positive for rabies diagnosis; 89% (312/351) of these occurred in urban areas and 85% (266/312) of the urban positive cases were among owned dogs. The mean age of the rabid dogs was 34 months and 61% were male. Aggressive behavior was observed in 77% of rabid dogs, followed by lack of coordination and paralysis (42%) and 48% of these dogs were responsible biting people or other animals. Information about vaccination status was obtained from 182 records and 51% of rabid dogs were non-vaccinated. The number of unvaccinated rabid dogs indicates a low vaccination index and this factor added to the high dog/man ratio must have contributed to the canine rabies epizootic observed in the studied area.

Filling of extraction sockets of feline maxillary canine teeth with autogenous bone or bioactive glass.

PURPOSE: To evaluate bone healing in the extraction socket of the feline maxillary canine tooth after grafting. METHODS: Eighteen adult cats were submitted to unilateral extraction of maxillary canine tooth and divided into three groups. In group 1 (n=6), control, the extraction socket was left empty. In group 2 (n=6), the extraction socket was filled with autogenous cancellous bone from the iliac crest and in group 3 (n=6), with bioactive glass particulate material. Cats were euthanized at four weeks postoperative. RESULTS: The radiographic examinations performed four weeks after surgery showed that in all groups the healing process converged to a radiopacity similar to that observed in the surrounding bones. Histological examination showed formation of woven bone within the extraction socket. The percentage of newly formed bone within the extraction socket, measured by the histometry, showed no statistically significant difference among the values of the three groups (Kruskal-Wallis'test p>0.05) (group 1: 63.96 ± 5.85, group 2: 66.84 ± 11.67, group 3: 59.28 ± 15.50). CONCLUSION: The bone regeneration observed in the extraction sockets filled with autogenous cancellous bone or bioactive glass was similar to that observed in the control sites, given an observation period of four weeks after extraction of the maxillary canine tooth.

Alveolar bone healing process in spontaneously hypertensive rats (SHR). A radiographic densitometry study.

UNLABELLED: Hypertension is one of the most important public health problems worldwide. If undiagnosed or untreated, this pathology represents a systemic risk factor and offers unfavorable conditions for dental treatments, especially those requiring bone healing. OBJECTIVES: The purpose of this study was to demonstrate, by analysis of bone mineral density (BMD), that the alveolar bone healing process is altered in spontaneously hypertensive rats (SHRs). MATERIAL AND METHODS: Wistar rats and SHRs were submitted to extraction of the upper right incisor and were euthanized 7, 14, 21, 28 and 42 days after surgery. Right maxillae were collected, radiographed and analyzed using Digora software. BMD was expressed as minimum (min), middle (med) and maximum (max) in the medium (MT) and apical (AT) thirds of the dental alveolus. RESULTS: The results were compared across days and groups. Wistar showed difference in med and max BMD in the MT between 7 and 28 and also between 14 and 28 days. The AT exhibited significant difference in med and min BMD between 7 and 28 days, as well as difference in min BMD between 28 and 42 days. SHRs showed lower med BMD in the MT at 28 days when compared to 21 and 42 days. Differences were observed across groups in med and min BMD at day 28 in the MT and AT; and in max BMD at 14, 21 and 42 days in the MT. CONCLUSIONS: These results suggest that the alveolar bone healing process is delayed in SHRs comparing with Wistar rats.

Copaiba oil in experimental wound healing in horses / Óleo de Copaíba na cicatrização de feridas experimentais em cavalos

ABSTRACT: The aim of this study was to evaluate the effects of 10% copaiba oil in experimentally induced wounds in horses. Four wounds were made in the lumbar and metacarpal regions of eight adult horses. In the treatment group, the wounds received 10% copaiba oil and in the control group 0.9% sodium chloride, in the daily dressing for 21 days. The wounds were evaluated three, 7, 14, and 21 days postoperatively. No significant differences were observed between the groups. The mean lumbar wound contraction rates were 80.54% and 69.64%, for the control and treated groups, respectively. For the wounds in the metacarpal region, these averages were 44.15% and 52.48%, respectively. Under the experimental conditions of the present study, it is concluded that 10% copaiba oil has beneficial in wound healing in the equine species and suggest that copaiba oil can be used as a therapeutic possibility in equine wound therapy.

RESUMO: O objetivo deste estudo foi avaliar os efeitos do óleo de copaíba a 10% em feridas induzidas experimentalmente em equinos. Quatro feridas foram confeccionadas nas regiões lombares e metacarpianas de oito cavalos adultos. No grupo tratamento, as feridas receberam óleo de copaíba a 10% e no grupo controle, cloreto de sódio a 0,9% nos curativos diários durante 21 dias. As avaliações das feridas foram feitas aos 3, 7, 14, e 21 dias de pós-operatório. Não foram observadas diferenças significativas entre os grupos estudados. As médias das taxas de contração das feridas lombares foram 79,24% e 69,64%, para o grupo controle e tratado, respectivamente. Nas feridas do metacarpo, estas médias foram 44,78% e 52,05%, respectivamente. Nas condições experimentais deste estudo, conclui-se que o óleo de copaíba a 10% foi benéfico na cicatrização de feridas na espécie equina, sendo uma alternativa terapêutica para cura de feridas nesta espécie.

Cryptosporidium spp. infection in mares and foals of the northwest region of São Paulo State, Brazil.

The present study aimed to analyze the occurrence of infection by Cryptosporidium spp. in mares and their respective foals. This study was carried out in 11 farms located in the municipalities of Araçatuba, Birigui, Guararapes and Santo Antônio do Aracangua, in the northwest region of the State of Sao Paulo, from November 2010 to March 2011. A total of 98 mares and 98 foals of several breeds were analyzed; among foals, 59 were males and 39 females, aged from three to 330 days. Feces were collected directly from the rectal ampulla, purified and processed according to modified Kinyoun stain. Occurrence of Cryptosporidium spp. was 21.4% (21/98) for foals and 18.4% (18/98) for mares. Occurrence of Cryptosporidium spp. had significant association with breeds and age of animals. Results obtained led to the conclusion that foals older than two months and Mangalarga animals are less susceptible to the occurrence of Cryptosporidium spp.

Salivary alpha amylase and cortisol levels in children with global developmental delay and their relation with the expectation of dental care and behavior during the intervention.

The purpose of this study was to analyze the alpha-amylase (sAA) and cortisol levels in children with Global developmental delay (GDD) before and after dental treatment and its association with the children's behavior during treatment. The morning salivary cortisol levels and activity of sAA of 33 children with GDD were evaluated before and after dental treatment and were compared to 19 healthy children. The behavior of children with GDD during dental care was assessed by the Frankl scale. Children with GDD showed lower levels of sAA activity than healthy children, but this result was not significant. The salivary cortisol levels were similar between GDD and healthy children. GDD children showed increased levels of sAA (but not cortisol) prior to the dental treatment as compared to the post-treatment phase. GDD children who showed less favorable behavior during dental care had higher levels of sAA and salivary cortisol than GDD children with more favorable behavior, but only the sAA results were significant. In conclusion, GDD children show hyperactivity of the SNS-axis in anticipation of dental treatment which indicates the need for strategies to reduce their anxiety levels before and during dental care.