Frequência de anticorpos anti-Leptospira spp. Em cães atendidos no hospital veterinário da Universidade Estadual Do Norte Do Paraná - UENP / Frequency of antibodies anti- Leptospira spp. In dogs treated at the veterinary hospital of Northern Paraná State University - UENP / Frecuencia de anticuerpos anti- Leptospira spp. En perros tratados en el hospital veterinario de la Universidad Estadual Del Norte De Paraná - UENP

A leptospirose é uma enfermidade bacteriana causada por espécies patogênicas do gênero Leptospira. Os roedores são considerados como principal reservatório, mas o cão, assim como outros animais domésticos, pode desenvolver a doença e se tornar carreadores assintomáticos. O objetivo desse trabalho foi conhecer a frequência de aglutininas anti-Leptospira spp. em cães atendidos no Hospital Veterinário da Universidade Estadual do Norte do Paraná (UENP), no município de Bandeirantes-Paraná. Foram testados, pela prova de soroaglutinação microscópica (SAM), 70 cães adultos assintomáticos para leptospirose, 27 machos e 43 fêmeas, de diferentes raças e idades. A frequência de animais positivos foi de 26,47%, 33,33% dos machos e 25,58% das fêmeas. Os sorovares Canicola (33,33%) e Autumnalis (27,78%) foram os mais frequentes, seguidos pelo Icterohaemorrhagiae (11%) e Grippotyphosa (11%). Conclui-se que a bactéria Leptospira está circulante em animais assintomáticos, podendo dispersar a doença para outros animais e inclusive para o homem...

Leptospirosis is a bacterial disease caused by pathogenic species of Leptospira. Rodents are considered the main reservoir, but dogs and other domestic animals may develop the disease and become asymptomatic carriers. The aim of this study was to determine the frequency of anti-Leptospira spp. agglutinins in dogs examined at the Veterinary Hospital of the Northern Paraná State University in Bandeirantes, Paraná, Brazil. A total of 70 asymptomatic adult dogs (27 male and 43 female), of different breeds and ages, were tested for leptospirosis by microscopic agglutination test. The frequency of positive animals was 26.47%, with 33.33% males and 25.58% females. Canicola (33.33%) and Autumnalis (27.78%) were the most frequent serovars, followed by Icterohaemorrhagiae (11%) and Grippotyphosa (11%). The authors concluded that the Leptospira bacterium is circulating in asymptomatic animals and can spread the disease to other animals and even to human beings...

La leptospirosis es una enfermedad bacteriana causada por especies patógenas del género Leptospira. Los roedores son considerados como reservorio principal, pero en el perro, así como otros animales domésticos, la enfermedad puede desarrollarse y convertirse en portadores asintomáticos. El objetivo de este estudio ha sido determinar la frecuencia de aglutininas anti-Leptospira spp. en perros atendidos en el Hospital Veterinario de la Universidad Estadual del Norte de Paraná (UENP), en el municipio de Bandeirantes-Paraná. Se realizó la prueba de suero aglutinación microscópica (SAM), en 70 perros adultos asintomáticos para leptospirosis, 27 machos y 43 hembras, de diferentes razas y edades. La frecuencia de animales positivos fue de 26,47%, 33,33% machos y 25,58% hembras. Los serovares Canicola (33,33%) y Autumnalis (27.78%) fueron los más frecuentes, seguidos por Icterohaemorrhagiae (11%) y Grippotyphosa (11%). Llegamos a la conclusión de que la bacteria Leptospira está circulante en animales asintomáticos, pudiendo dispersar la enfermedad a otros animales e incluso a los seres humanos...

Zoonoses in humans from small rural properties in Jataizinho, Parana, Brazil.

The aim of this study was to conduct a serological survey for Lyme diseases, brucellosis, leptospirosis and toxoplasmosis and identify the risk variables related to these zoonoses in humans living in the rural area of Jataizinho, state of Parana, Brazil. A total of 63 rural properties were surveyed. Additionally, 207 serum samples collected from these rural area inhabitants were tested for indirect immunofluorescence (IFI) and western blots (WB) were performed to detect Borrelia burgdorferi (sensu lato); a tamponated acidified antigen test (AAT) and 2-mercaptoethanol (2-ME) were used to detect antibodies of Brucella abortus; the microscopic agglutination test (MAT) was carried out to detect antibodies anti-Leptospira spp. and IFI was used to find antibodies of Toxoplasma gondii. Two of the samples (0.96%) were reactive for Lyme borreliosis, three (1.4%) for brucellosis, 25 (12.1%) for leptospirosis and 143 (69.1%) for toxoplasmosis. Although the town of Jataizinho has a human development index (IDH) that was considered to be average (0.733) in the state of Parana, the low social, economic and cultural conditions of the population from small rural properties have resulted in lack of basic information on animal health and direct or indirect contact with the various species of domestic animals, wildlife and ticks have probably contributed to the prevalence levels found. These results show the need for additional regional studies in order to determine the epidemiological characteristics of these diseases as well as their respective vectors and reservoirs so that effective prophylaxis can be administered in the human population.

Toxoplasmosis, leptospirosis and brucellosis in stray dogs housed at the shelter in Umuarama municipality, Paraná, Brazil.

BACKGROUND: Leptospirosis, toxoplasmosis and brucellosis are diseases with worldwide distribution. Among stray dogs, these zoonoses are facilitated by direct contact with other animal species, by the habit of scavenging garbage and hunting in search of food, drinking standing water, smelling other animals' urine, licking female genitalia and the sexual act itself. The objective of this study was to detect antibodies anti-Toxoplasma gondii, anti-Leptospira spp., anti-Brucella canis and anti-Brucella abortus in stray dogs housed in shelters at Umuarama city, Paraná, Brazil. In order to detect toxoplasmosis, indirect immunofluorescence assay (IFA) was performed, agglutination microscopic (MAT) test for leptospirosis and agar gel immunodiffusion (AGID) and buffered acidified antigen (BAA) tests for brucellosis. RESULTS: Of the 175 serum samples analyzed, 70.85% were considered positive for toxoplasmosis by IFA, 20% by MAT for leptospirosis and 2.85% by AGID for Brucella canis. CONCLUSIONS: The serological results of this study showed that stray dogs housed at the private shelter are potential carriers of these three different zoonoses and contribute to the spread and maintenance of these etiologic agents in the urban area of Umuarama (PR), Brazil.

First record of Borrelia burgdorferi B31 strain in Dermacentor nitens ticks in the northern region of Parana (Brazil).

The aim of this study was to investigate the presence of DNA of Borrelia burgdorferi sensu lato (s.l.) in ticks that feed on horses used for animal traction in rural Jataizinho, Parana, Brazil. Between February and June 2008, a total of 224 ticks was collected of which 75% were identified as Dermacentor nitens and 25% as Amblyomma cajenense. To amplify B. burgdorferi s.l. DNA, the intergenic space region (ISR) between the 5S (rrf) 23S (rrl) rRNA genes was used as targets for nested-PCR. Two ticks of the D. nitens species were positive for B. burgdorferi s.l. Both species showed a fragment of 184 bp, but the sequencing revealed 99.9% homology with the B. burgdorferi sensu stricto (s.s.) strain B31. These results showed, for the first time, the presence of spirochete DNA infecting ticks that parasitize horses used for animal traction, in the rural municipality mentioned. In conclusion, this study opens up promising prospects for determining the infection rate of B. burgdorferi s.s. genospecies or other species in the equine population, as well as the impact of the infection rate on Lyme disease in the state of Parana.

Occurrence of Ehrlichia canis and Anaplasma platys in household dogs from northern Parana.

Canine monocytic ehrlichiosis caused primarily by Ehrlichia canis and canine thrombocytic anaplasmosis induced by Anaplasma platys are important emerging zoonotic tick-borne diseases of dogs. There is evidence that these pathogens can also affect humans. This study evaluated the presence of E. canis and A. platys in blood samples collected from 256 domiciled dogs in the municipality of Jataizinho, located in north region of the State of Parana, Brazil, by PCR assay. The occurrence of E. canis and A. platys was 16.4% (42/256) and 19.4% (49/256), respectively; while 5.47% (14/256) of the dogs evaluated were co-infected by these two organisms. The presence of E. canis and A. platys was not significantly associated with the variables evaluated (sex, age, outdoor access, and presence of ticks during blood collection). Infection of dogs by E. canis was associated with anemia and thrombocytopenia, while infection induced by A. platys was related only to thrombocytopenia. Canine monocytic ehrlichiosis and canine thrombocytic anaplasmosis should be included in the differential diagnoses when these hematological alterations are observed during routine laboratory evaluation of dogs.

Blockade of irradiation-induced autophagosome formation impairs proliferation but does not enhance cell death in HCT-116 human colorectal carcinoma cells.

This work was undertaken to gain further information on the molecular mechanisms underlying autophagosome formation and its relation with tumor cell survival in response to radiation in colon cancer. A human colon cancer cell line, HCT-116, was examined with respect to cell survival after blockade of irradiation-induced autophagosome formation by pharmacological interference. Autophagosome formation was confirmed using a kinetic study with incorporated bovine serum albumin gold-conjugate (BSA-Au) analyzed by electron microscopy and an autophagosome-associated LC3B antibody measured by immunofluorescence and Western blotting. Annexin V/PI double staining was used to monitor cell death by apoptosis, and cell cycle profiles by flow cytometry. Ionizing radiation (IR) promoted autophagosome formation in the HCT-116 IR-surviving cells. Pharmacological interference showed that PI3K/Akt and Src were involved in early stages of autophagosome formation. IR alone decreased cell proliferation by arresting cells in the G2/M phase, and pharmacological interference of autophagosome formation decreased proliferation, but did not affect cell survival. Also, our data suggest that decreased proliferation caused by PI3K and Src inhibitors could be through S phase cell cycle delay. Our results clearly indicate that blockade of IR-induced autophagosome formation impairs proliferation but does not enhance cell death in colon cancer cells.

Prevalence of Leptospira interrogans antibodies in free-ranging Tayassu pecari of the Southern Pantanal, Brazil, an ecosystem where wildlife and cattle interact.

We surveyed a wild population of white-lipped peccaries (Tayassu pecari) in the Brazilian Pantanal for evidence of Leptospira interrogans. Serum samples from 71 free-ranging T. pecari were obtained between 2003 and 2005 in the southern Pantanal of Mato Grosso do Sul state. We used microscopic microagglutination to test for antibodies against 14 L. interrogans serovars (antibody titers ≥ 1:100 were considered seropositive). Seventy percent of captured animals tested positive for leptospirosis antibodies. Antibodies against icterohaemorrhagiae and autumnalis serovars were the most prevalent. We used log-linear analyses to test for associations among seropositivity, age class, and sex of captured animals. Seropositivity was strongly associated with animal age class, but independent of sex. Forty-six percent of animals less than 2 years old, 63% of adults during peak reproductive years, and 100% of the oldest age class were seropositive. A nonparametric multivariate procedure (MRPP) showed that the composition of serovar antibody types changed with age, and ANOVA models demonstrated that antibody titers increased with age, suggesting long-term exposure to a greater number and variety (i.e., serovar types) of L. interrogans infections. This study presents the first quantitative survey of antibodies against L. interrogans serovars in a T. pecari population of the Pantanal. The high prevalence of leptospirosis antibodies in free-ranging white-lipped peccaries and the potential impacts on reproduction and population dynamics emphasize the need for further studies investigating the roles of Pantanal wildlife and livestock in the transmission and maintenance of L. interrogans in the environment.

PI3K/Akt and GSK-3ß prevents in a differential fashion the malignant phenotype of colorectal cancer cells.

PURPOSE: During colorectal cancer progression, the loss of differentiation and cell-cell adhesion as well as a higher migratory potential are well-defined features; however, the signaling mechanism governing these events is not fully elucidated. The aim of this study was to investigate the role that PI3K and downstream effectors play in controlling colon cancer malignant phenotypes. METHODS: HCT-116 cells, a human model of colon cancer, which are highly metastatic and undifferentiated, were treated with LY294002, a specific inhibitor of PI3K. Cell differentiation and apical junctional complex (AJC) formation were monitored using alkaline phosphatase and electron microscopy analysis. Immunofluorescence and Western blotting were used to accompany the subcellular localization of AJC proteins. PI3K downstream molecules were analyzed by western blotting, and proliferation, wound healing, and colony formation techniques to determine malignant phenotype alterations. RESULTS: PI3K inhibition increased alkaline phosphatase activity, led to an enterocyte-like growth and formed a functional AJC. LY294002 treatment was able to recruit E-cadherin, ß-catenin, claudin-3, and ZO-1 to the cell-cell contact region, and this effect was essential for AJC assembly and association of these proteins to the cytoskeleton. Furthermore, we provided evidence that PI3K inhibition leads to a decrease in p-Akt and p-GSK-3ß and increased p-ß-catenin levels, which in turn controlled cell proliferation, motility, and colony formation. CONCLUSION: Our results demonstrate that PI3K/Akt and GSK-3ß prevents in a differential fashion the malignant phenotype of HCT-116 colorectal cancer cells, which could constitute a potential therapeutic target for treatment of this cancer type.

Safety of general anesthesia for lumbar puncture and bone marrow aspirate/biopsy in pediatric oncology patients.

INTRODUCTION: Painful short duration procedures such as bone marrow aspiration/biopsy and the lumbar puncture with or without intrathecal chemotherapy are frequently performed during the treatment of children with cancer. This study evaluated the frequency and severity of complications of bone marrow aspiration biopsy and lumbar puncture/intrathecal chemotherapy under general anesthesia. PATIENTS AND METHODS: A prospective observational study was performed from November 2003 to August 2005. Patients with cancer younger than 21 years old, receiving treatment at the Pediatric Oncology Unity of Hospital de Clínicas de Porto Alegre, undergoing diagnostic and/or therapeutic short duration procedures carried out under general anesthesia in the outpatient surgery unit. RESULTS: One hundred and thirty-seven patients were submitted to 423 procedures under general anesthesia. There were 61% boys, mean age of 7.5 years (0.2 to 21) and ASA II 98%. Eighty seven percent of the procedures were carried out in patients with leukemia or lymphoma. The majority of the procedures had no adverse events during intraoperative and postoperative periods. No procedure had to be suspended after it had begun. One patient had lumbar pain after the procedure and was admitted to the ward with suspected subdural bleeding, but this was not confirmed. No patient needed cardiopulmonary reanimation or treatment in the intensive care unit. CONCLUSIONS: General anesthesia for short duration painful procedures in children undergoing treatment for malignancies is safe when carried out by trained professionals in outpatient clinical surgery unit.

Presence of domesticated cats and visual impairment associated to Toxoplasma gondii serum positive children at an elementary school in Jataizinho, state of Paraná, Brazil.

Toxoplasmosis possess worldwide geographic distribution and high serological prevalence in human beings and animals. Acquired way is, in the majority of the times, light or asymptomatic however, in approximately 15% of the cases can be developed the ocular form. The aim of this study is to research the occurrence of IgG anti- Toxoplasma gondii antibodies in children at an elementary school in Jataizinho (PR) and relate some factors with the epidemiology of the toxoplasmosis. A total of 276 samples of blood by using indirect immunofluorescence (IFA) where 128 (46.4%) were detected positives. The analysis of the variables indicated that the presence of domestic cats in the household was an important factor associated to the infection by T. gondii (OR= 3.45; 1.61- 7.45) as well as to the children who described any kind of visual impairment (OR= 3.19; 1.11 - 9.35).

Preoperative intravenous clonidine in the surgical treatment of cataract: evaluation of the clinical benefits.

BACKGROUND AND OBJECTIVES: Clonidine has been used in anesthesia for its sedative and analgesic properties, and to achieve greater hemodynamic stability with reduction in intra-ocular pressure. This study evaluated the clinical effects of intravenous clonidine (2.5 microg.kg(-1)) on the intra-ocular pressure (IOP), hemodynamic parameters, and post-anesthetic recovery in the surgical treatment of cataract. METHODS: This is a randomized, double-blind, placebo-controlled study that included patients undergoing cataract surgery under peribulbar block. After placement of monitoring devices, baseline (M0) values of IOP, mean arterial pressure (MAP), heart rate (HR), SpO2, and the degree of sedation (Ramsay) were determined. Afterwards, placebo (Group P) or clonidine (Group C) was administered intravenously. After 30 minutes, at Moment 1 (M1), measurements were repeated, and once again at the end of the surgery, at Moment 2 (M2). RESULTS: The change in IOP between M0 and M1 was different for both groups, 14.5% in Group C and 5.25% in Group P (p = 0.01), and the means remained reduced until M2. Mean arterial pressure in Group C showed a 17% reduction from M0 to M1 and increased 3.5% in Group P (p < 0.001) and in Group C it showed a recovery at M2 (p = 0.17). The heart rate decreased 6.4% in Group C and 1% in Group P (p = 0.1) from M0 to M1. There was a greater sedation in Group C when compared with Group P (p < 0.001) but there was no cases of respiratory depression. The relative risk of intraoperative hypertension was 0.657 (CI 95% 0.517 to 0.835) in Group C, p < 0.01. Differences in the incidence of tachycardia, hypoxemia, hypotension, and increased hospital staying were not observed between both groups. CONCLUSIONS: Clonidine, under the conditions of the present study, showed to be a safe drug and decreased IOP and the intraoperative risk of hypertension without delaying hospital discharge.

Isolation of leptospira Serovars Canicola and Copenhageni from cattle urine in the state of ParanÁ, Brazil.

In 2001, 698 urine samples were randomly collected from cattle at a slaughterhouse in the State of Paraná, Brazil. Direct examination using dark field microscopy was carried out immediately after collection. Five putative positive samples were cultured in modified EMJH medium, yielding two positive cultures (LO-14 and LO-10). Typing with monoclonal antibodies revealed that the two isolates were similar to Canicola (LO-14) and Copenhageni (LO-10). Microscopic agglutination test results show that Hardjo is the most common serovar in cattle in Brazil. Rats and dogs are the common maintenance hosts of serovars Copenhageni and Canicola. The excretion of highly pathogenic serovars such as Copenhageni and Canicola by cattle can represent an increasing risk for severe leptospirosis is large populations, mainly living in rural areas.

Seroepidemiology and occupational and environmental variables for leptospirosis, brucellosis and toxoplasmosis in slaughterhouse workers in the Paraná State, Brazil.

Leptospirosis, brucellosis and toxoplasmosis are widely-distributed zoonosis, being the man an accidental participant of their epidemiological chains. The aim of this paper was to make a seroepidemiological report and identify occupational and environmental variables related to these illnesses in 150 workers in a slaughterhouse in the Northern region of Paraná. For the diagnosis of leptospirosis a microscopical seroagglutination test was applied; for brucellosis, the tamponated acidified antigen test and the 2-mercaptoetanol tests were used, and for toxoplasmosis the indirect immunofluorescence reaction test. For each employee an epidemiological survey was filled, which investigated occupational and environmental variables which could be associated with these infections. Positive results for leptospirosis were found in 4.00% of the samples, for brucellosis in 0.66% of samples and toxoplasmosis in 70.00%. From the three diseases researched, only the results for leptospirosis suggest occupational infection.