The platform switching concept revisited. An experimental study in dogs

Objective: To evaluate the influence of the configuration of the marginal aspect of implants placed immediately into extraction sockets on peri-implant hard tissue adaptation. Material and methods: In 6 Labrador dogs, endodontic treatments of the mesial roots of 1M1 were performed and the distal roots were removed. 2P2 was extracted as well. Implants were immediately placed in the center of the distal alveoli. Cylindrical straight implants were installed in the right side of the mandible (Control), while, in the left side, implants with a reduced diameter in the coronal portion, yielding an indentation in the surface continuity (Test), were installed. Cover screws were affixed, and the flaps were sutured to allow non-submerged healing. After 4 months of healing, histological slides were obtained for assessments. Results: A buccal resorption of 1.58 ± 1.28 and 1.90 ± 1.93 mm at the control and of 0.26 ± 0.90 and 0.14 ± 0.66 mm at the test sites was observed at the premolar and molar regions, respectively. The buccal coronal level of osseointegration was located apically to the margin of the smooth/rough surface border by 2.40 ± 0.90 and 3.70 ± 0.87 mm at the control sites and 1.19 ± 0.45 and 2.16 ± 0.96 mm at the test sites at the premolar and molar sites, respectively. All differences yielded statistical significance. Conclusions: The use of implants with a reduced diameter in their coronal aspect may contribute to preservation of the buccal bony crest in a more coronal level compared with conventional implants. Thus, the study confirmed the efficacy of the platform switching concept. © 2013 John Wiley & Sons A/S.

Asymptomatic dogs are highly competent to transmit Leishmania (Leishmania) infantum chagasi to the natural vector

We evaluated the ability of dogs naturally infected with Leishmania (Leishmania) infantum chagasi to transfer the parasite to the vector and the factors associated with transmission. Thirty-eight infected dogs were confirmed to be infected by direct observation of Leishmania in lymph node smears. Dogs were grouped according to external clinical signs and laboratory data into symptomatic (n= 24) and asymptomatic (n= 14) animals. All dogs were sedated and submitted to xenodiagnosis with F1-laboratory-reared Lutzomyia longipalpis. After blood digestion, sand flies were dissected and examined for the presence of promastigotes. Following canine euthanasia, fragments of skin, lymph nodes, and spleen were collected and processed using immunohistochemistry to evaluate tissue parasitism. Specific antibodies were detected using an enzyme-linked immunosorbent assay. Antibody levels were found to be higher in symptomatic dogs compared to asymptomatic dogs (p= 0.0396). Both groups presented amastigotes in lymph nodes, while skin parasitism was observed in only 58.3% of symptomatic and in 35.7% of asymptomatic dogs. Parasites were visualized in the spleens of 66.7% and 71.4% of symptomatic and asymptomatic dogs, respectively. Parasite load varied from mild to intense, and was not significantly different between groups. All asymptomatic dogs except for one (93%) were competent to transmit Leishmania to the vector, including eight (61.5%) without skin parasitism. Sixteen symptomatic animals (67%) infected sand flies; six (37.5%) showed no amastigotes in the skin. Skin parasitism was not crucial for the ability to infect Lutzomyia longipalpis but the presence of Leishmania in lymph nodes was significantly related to a positive xenodiagnosis. Additionally, a higher proportion of infected vectors that fed on asymptomatic dogs was observed (p= 0.0494). Clinical severity was inversely correlated with the infection rate of sand flies (p= 0.027) and was directly correlated with antibody levels (p= 0.0379). Age and gender did not influence the transmissibility. Our data show that asymptomatic dogs are highly infective and competent for establishing sand fly infection, indicating their role in maintaining L. (L.) infantum chagasi cycle as well as their involvement in VL spreading in endemic areas. © 2013 Elsevier B.V.

Leishmaniasis causes oxidative stress and alteration of oxidative metabolism and viability of neutrophils in dogs

The aim of the present study was to test the hypothesis that oxidative stress and alteration of oxidative metabolism and apoptosis of neutrophils in dogs vary with the stage of leishmaniasis and to determine the contribution of uremia to such alterations. Dogs with leishmaniasis were classified into two stages: moderate (Leish II, n = 20) or very severe (i.e. with concurrent uremia; Leish IV, n = 20) according to the LeishVet Consensus. The two leishmaniasis groups were compared with uremic dogs without leishmaniasis (Uremic, n = 10) and to healthy dogs (Control, n = 30). To determine oxidative stress, total antioxidant/oxidant capacity, lipid peroxidation, total glutathione and the plasma antioxidants albumin, uric acid and bilirubin were quantified. Superoxide production was determined using the hydroethidine probe and viability and apoptosis were measured using annexin V-PE by capillary flow cytometry. Oxidative stress was present in both uremia and leishmaniasis with reduced total antioxidant capacity and was associated with increased induced production of superoxide and apoptosis. The greatest amount of oxidants was observed in animals with moderate disease only. Neutrophils from uremic dogs with and without leishmaniasis had decreased viability and an increased apoptosis rate in addition to increased lipid peroxidation. In conclusion, oxidative stress occurs in both stages of leishmaniasis with differences in intensity and levels of plasma markers; however, uremia does contribute to the decreased spontaneous viability of neutrophils in dogs in the final stage of the disease. © 2013 Elsevier Ltd. All rights reserved.

CD95 (FAS) and CD178 (FASL) induce the apoptosis of CD4+ and CD8+ cells isolated from the peripheral blood and spleen of dogs naturally infected with Leishmania spp

Infected dogs are urban reservoirs of Leishmania chagasi, which is a causative agent of visceral leishmaniasis (VL). Dogs exhibit immune suppression during the course of this disease, and lymphocyte apoptosis is involved in this process. To investigate apoptosis and the expression levels of FAS-FAS-associated death domain protein (CD95 or APO-1), FASL-FAS ligand protein (CD178), and TRAIL-TNF-related apoptosis-inducing ligand (CD253) receptors in peripheral blood mononuclear cells and spleen leukocytes from 38 symptomatic dogs with moderate VL and 25 healthy dogs were evaluated by flow cytometry. The apoptosis rate of blood and splenic CD4+ and CD8+ cells was higher in infected dogs than in healthy dogs. The expression levels of FAS and FASL in blood and splenic CD4+ cells were lower in infected dogs than in healthy dogs. FAS expression in CD8+ cells was higher in infected dogs than in healthy dogs; in contrast, FASL expression was lower in infected dogs. The expression of the TRAIL receptor increased only in splenic CD8+ cells from infected dogs. The FAS and FAS-L blocking antibodies confirmed the importance of these receptors in apoptosis. Our results enhance the current understanding of the immune response in dogs infected with L. chagasi, facilitating the future development of therapeutic interventions to reduce lymphocyte depletion. © 2013 Elsevier B.V.

Longitudinal analysis of serological tests officially adopted by the Brazilian Ministry of Health for the diagnosis of canine visceral leishmaniasis in dogs vaccinated with Leishmune®

Development of vaccines against canine visceral leishmaniasis (CVL) may provide a prophylactic barrier, but antibody response detected by standard diagnostic techniques may not separate vaccinated from naturally infected dogs. Moreover, anti-Leishmania antibody levels in vaccinated dogs may be detectable for months. Accordingly, the aim of the present study was to comparatively evaluate an in-house ELISA with three serological tests officially adopted by the Brazilian Ministry of Health for the diagnosis of CVL in dogs vaccinated with Leishmune®. A total of 18 mongrel dogs were submitted to a complete protocol of the vaccine, monitored and evaluated in 5 times (T0-T4) up to 180 days after T0. Twenty-one days after the first dose (T1), 50% of the dogs were seropositive by the in-house ELISA and 5.5% by IFAT, while by the official ELISA and DPP® CVL rapid test all dogs tested negative. At time T2, 42 days after of the first dose, 100%, 83.3%, 11.1%, and 5.5% of the dogs were seropositive by the in-house ELISA, IFAT, official ELISA kit and the DPP® CVL rapid test, respectively. Ninety days after the first dose (T3), 100%, 83.3%, 72.2% and 33.3% of the dogs were seropositive by the in-house ELISA, official ELISA kit, IFAT, and the DPP® CVL rapid test, respectively. Finally, at time T4, 88.8%, 33.3%, 11.1% and 5.5% of the dogs were seropositive by the in-house ELISA, official ELISA kit, DPP® CVL rapid test and IFAT, respectively. In conclusion, dogs vaccinated with Leishmune® cross-react by an in-house ELISA and by the three official Brazilian serological tests for the diagnosis of canine visceral leishmaniasis up to six months after the first vaccine dose, and may be mistakenly diagnosed and removed. © 2013 Elsevier B.V.

Canine visceral leishmaniasis due to Leishmania (L.) infantum chagasi in Amazonian Brazil: comparison of the parasite density from the skin, lymph node and visceral tissues between symptomatic and asymptomatic, seropositive dogs

A leishmaniose visceral canina (LVC) é reconhecida pelas características clínicas da doença e é altamente letal. A infecção, entretanto, pode ser totalmente assintomática em alguns cães soropositivos, o que tem levantado questão polêmica sobre a possibilidade desses animais, serem ou não uma fonte importante da infecção para o flebotomíneo, Lutzomyia longipalpis, o principal vetor da leishmaniose visceral americana (LVA). Neste estudo foram examinados 51 cães com LVC aguda, provenientes de área endêmica de LVA no Estado do Pará, Brasil, e a carga parasitária, formas amastigotas de, na pele, linfonodo poplíteo e vísceras (fígado e baço) foi comparada com a de nove cães assintomáticos soropositivos (IFAT-IgG). Fragmentos de biópsia desses tecidos obtidos post-mortem foram processados para análise através de imunohistoquímica, usando um anticorpo policlonal contra Leishmania sp. Os testes do Qui-quadrado (X²) e Mann Whitney foram usados para avaliar as médias da densidade de macrófagos infectados (p < 0,05). Os resultados mostraram que não houve diferença (p > 0,05) na densidade de macrófagos infectados da pele (10,7/mm² x 15,5/mm²) e do linfonodo (6,3/mm² x 8,3/mm²) entre cães assintomáticos e sintomáticos. Entretanto, a densidade de macrófagos infectados da víscera de cães sintomáticos (5,3/mm²) foi maior (p < 0,05) que a de cães assintomáticos (1,4/mm²). Estes resultados sugerem, fortemente, que cães naturalmente infectados por L. (L.) i. chagasi, assintomáticos ou sintomáticos, podem servir como fonte de infecção, principalmente, considerando-se que a densidade de macrófagos infectados da pele (10,7/mm² x 15,5/mm²), local onde o flebotomíneo vetor Lu. longipalpis realiza a hematofagia, foi maior (p < 0,05) que as do linfonodo (6,3/mm² x 8.3/mm²) e vísceras (1,4/mm2x 5,3/mm2).

Serological cross-reactivity of Trypanosoma cruzi, Ehrlichia canis, Toxoplasma gondii, Neospora caninum and Babesia canis to Leishmania infantum chagasi tests in dogs

Introduction: The aim of this study was to evaluate the serological cross-reactivity between Leishmania sp. and other canine pathogens. Methods: Positive serum samples for Ehrlichia canis, Babesia canis, Toxoplasma gondii, Neospora caninum and Trypanosoma cruzi were tested using three serological methods enzyme linked immunosorbent assay (ELISA), indirect immunofluorescent antibody test (IFAT) and Kalazar Detect™, for canine visceral leishmaniasis. Results: Of the 57 dog samples tested, 24 (42.1%) tested positive using one of the three serological methods: 10/57 (17.5%) for ELISA, 11/57 (19.3%) for IFAT and 3/57 (5.3%) for Kalazar Detect™. Conclusions: Our results demonstrated that the presence of other infectious agents may lead to cross-reactivity on leishmaniasis serological tests.

Hemodynamic and antiarrhythmic effects of lidocaine or amiodarone in dogs anesthetized with halothane

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cardiovascular and respiratory alterations in dogs during cervical myelography using ioversol

Subarachnoid infusion of most contrast mediums and the steps involved in performing a cervical myelography have adverse affects that can discourage its use in the radiographic diagnosis of spinal cord diseases. Thus, the cardiovascular and respiratory alterations associated with neck flexion, subarachnoid puncture, and cerebrospinal fluid drainage during subarachnoid infusion of ioversol (320mgI/mL) in dogs under general anesthesia using isoflurane were evaluated. The dogs received subarachnoid infusion of autologous cerebrospinal fluid kept at 38°C - control group (GC); ioversol 0.3mL/kg at 25°C (GI25) and ioversol 0.3mL/kg heated to 38°C (GI38). Each dog had its heart rate (HR), systolic arterial pressure (SAP), diastolic arterial pressure (DAP), respiratory rate (RR), oxyhemoglobin saturation (SaO2) and electrocardiography readings (PR and QT intervals) recorded. Group comparisons showed no statistical difference regarding neck positioning, subarachnoid puncture, and subarachnoid infusion of contrast medium on HR, RR and SaO2, cardiac rhythm or conduction. However, isoflurane significantly increased PR and QT intervals. Based on these findings, it is concluded that the steps involved in cervical myelography and the use of ioversol 320mgI/mL at 0.3mL/kg (25ºC and 38ºC) during cervical myelography did not result in relevant cardiovascular and respiratory alterations, except for an elevation in arterial pressure after injection of ioversol.

Hematologic changes in propofol-anesthetized dogs with or without tramadol administration

Drugs commonly used in anesthesia practice may significantly alter the oxidative state of blood cells. This mechanism could contribute to the immune suppression that occurs transiently in the early postoperative period. Thus, we assessed the effects of continuous rate infusion (CRI) of propofol associated or not with tramadol on hematologic parameters in dogs. Eight adult mongrel dogs were anesthetized on 2 occasions, 15 d apart. Two groups were formed: control group (CG) and tramadol group (GT). Propofol was used for induction (10mg kg-1) followed by a CRI (0.7mg kg-1minute-1). The animals were positioned in lateral recumbency and mechanically ventilated with inspired oxygen fraction of 0.6. In TG, tramadol (2mg kg-1) followed by a CRI (0.5mg kg-1minute-1) was administered in dogs. In the CG the sodium chloride (NaCl) solution at 0.9% was administered followed by its CRI, in the same volume that was used in TG. The measurement was taken before anesthesia induction (Tbasal), 30 minutes after induction (T0) and then at 30-minute intervals (T30 to T60). Red blood cells, hematocrit, hemoblogin concentration and total leukocytes count decreased from T0 in both groups. In TG, lymphocytes count at Tbasal [1.86 (0.82) x103µl-1] was greater than at T0, T30 and T60 [0.96(0.50), 0.92(0.48) and 0.95(0.48) x103µl-1, respectively]. No significant differences were observed for platelets neutrophil, eosinophil, basophil and monocyte count. In dogs, propofol-anesthesia associated or not with tramadol promoted decrease in blood cell count and should be used with caution in immunossupressed patients.

Brainstem auditory evoked potential testing in Dalmatian dogs in Brazil

The brain stem auditory-evoked potential (BAEP) is an electrophysiologic test that detects and records the electrical activity in the auditory system from cochlea to midbrain, generated after an acoustic stimulus applied to the external ear. The aim of this study is to obtain normative data for BAEP in Dalmatian dogs in order to apply this to the evaluation of deafness and other neurologic disorders. BAEP were recorded from 30 Dalmatian dogs for a normative Brazilian study. Mean latencies for waves I, III, and V were 1.14 (±0.09), 2.62 (±0.10), and 3.46 (±0.14) ms, respectively. Mean inter-peak latencies for I-III, III-V, and I-V intervals were 1.48 (±0.17), 0.84 (±0.12), and 2.31 (±0.18) ms, respectively. Unilateral abnormalities were found in 16.7% of animals and bilateral deafness was seen in one dog. The normative data obtained in this paper is compatible with other published data. As far as we know this is the first report of deafness occurrence in Dalmatian dogs in Brazil.

Nonsteroidal anti-inflammatory therapy: changes on renal function of healthy dogs

PURPOSE: To evaluate the renal function in healthy dogs submitted to nonselective and preferential COX-2 nonsteroidal anti-inflammatory drug (NSAID) therapy. METHODS: Twenty four healthy dogs were distributed into four groups (G) (n=6): ketoprofenG - treated with ketoprofen; nimesulideG - treated with nimesulid; meloxicanG - treated with meloxican; and etodolacG - treated with etodolaco. All the dogs received the NSAIDs for 10 days by oral route. Physical examination and renal function (urinalysis, urinary sodium and gamma-glutamyl transpeptidase (GGT), serum urea, creatinine, potassium and sodium, and endogenous creatinine clearance) were evaluated before, after five and ten days (T0, T5 and T10) of the treatment in all groups. RESULTS: Changes were observed in urinalysis, with a significant increase in renal cells in the urine at T5 and T10 in nimesulideG. Significant reduction in urinary sodium in nimesulideG at T5 was observed. The clearance values were lower in ketoprofenG at T10. CONCLUSIONS: Meloxicam and etodolac were the drugs that have proven to be safer for short-term therapy in healthy dogs in relation to renal function. NSAIDs ketoprofen and nimesulide should be used judiciously in dogs with renal dysfunction, since there are promoted changes in renal function.

Molecular and serological detection of tick-borne pathogens in dogs from an area endemic for Leishmania infantum in Mato Grosso do Sul, Brazil

Patógenos transmitidos por carrapatos atingem uma variedade de hospedeiros vertebrados. Para identificar os agentes patogênicos transmitidos por carrapatos entre cães soropositivos para Leishmania infantum no município Campo Grande-MS, foi realizado um estudo sorológico e molecular para a detecção de Ehrlichia canis, Anaplasma platys e Babesia vogeli em 60 amostras de soro e baço, respectivamente. Adicionalmente, foi realizado o diagnóstico confirmatório de L. infantum por meio de técnicas sorológicas e moleculares. Também foi realizado o alinhamento e análise filogenética das sequências para indicar a identidade das espécies de parasitas que infectam esses animais. Anticorpos IgG anti-Ehrlichia spp., anti-B. vogeli e anti-L. infantum foram detectados em 39 (65%), 49 (81,6%) e 60 (100%) dos cães amostrados, respectivamente. Vinte e sete (45%), cinquenta e quatro (90%), cinquenta e três (88,3%), dois (3,3%) e um (1,6%) cães mostraram-se positivos na PCR para E. canis, Leishmania spp., Leishmania donovani complex, Babesia sp. e Anaplasma sp., respectivamente. Após o seqüenciamento, os amplicons mostraram 99% de similaridade com isolados de E. canis, B. vogeli e A. platys e Leishmania chagasi. Os resultados deste estudo indicaram que os cães soropositivos para L. infantum de Campo Grande, MS, são expostos a vários agentes transmitidos por carrapatos, e, portanto, devem ser incluídos no diagnóstico diferencial em cães com suspeita clínica de leishmaniose.

Prevalence of antibodies against Toxoplasma gondii and Neospora caninum in dogs

Toxoplasmosis and neosporosis are parasitic diseases of global importance. The present study had the objective to determine the influence of age, sex and breed in the prevalence of antibodies against both diseases in dogs from Brotas city, São Paulo State, Brazil. Blood samples of 342 dogs were collected, and the age, sex and breed of each animal were recorded. The serological diagnosis for toxoplasmosis and neosporosis were performed using the immunofluorescent antibody test (IFAT). The Fischer's test was used to calculate the association probability of the variables, with a = 5%. For toxoplasmosis the prevalence of antibodies was 26.9% (CI 95% 22.4-31.8%), and for neosporosis 4.97% (CI 95% 3.1-7.8%). The statistical analysis revealed a higher risk of infection for T. gondii in dogs with more than five years.

Non-contact specular microscopy in aphakic and pseudophakic dogs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)