Investigação soroepidemiológica de leptospirose canina na área territorial urbana de Botucatu, São Paulo, Brasil

Leptospirosis was seroepidemiologically investigated in 775 dogs from Botucatu, state of Sao Paulo, in blood samples collected during the annual anti-rabies vaccination campaign. The samples were collected from 14 out of the 45 vaccination stations geographically distributed in five different municipal districts, with a territorial urban area of 31Km2. Samples were taken from 449 males and 326 females; 564 were of undetermined breed, and 211 were pedigree animals; ages ranged from 3 months to 20 years. Diagnosis included the microscopic serum agglutination tests using 12 Leptospira spp serovars. Serological results and epidemiological variables were compared using Goodman and chi-square tests, with χ=0.05. There were 119 (15.3%) positive samples for 11 serovars; the most important was canicola with 48 (40.3%), followed by pyrogenes 41 (34.5%). Statistical analysis showed significant difference for: breed, 17.7% positive in undetermined breeds and 9.0% in pedigrees; and sex, 18.4% positive in males and 11.0% in females. There was no statistically significant difference in relation to age with uniform distribution of positives.

Estimativa da febre maculosa em cães para a vigilância e motitoramento da antropozoonose no municipio de Botucatu, no Estado de São Paulo

Pós-graduação em Medicina Veterinária - FMVZ

Efeito das suplementações de selênio na resposta imune humoral anti-rábica, na concentração sérica de selênio e do cortisol em bovinos

Pós-graduação em Medicina Veterinária - FMVZ

Proteção imunológica contra raiva em população rural exposta à epidemia em 2005

No mês de maio de 2005, no município de Augusto Correa, nordeste do Estado do Pará, foi diagnosticado um surto de raiva paralítica transmitida por morcegos hematófagos que resultou em 15 casos confirmados da doença. A população exposta recebeu vacinação anti-rábica em regime de pré e pós-exposição, sendo atendidas aproximadamente 3500 pessoas. O presente estudo verificou a persistência de anticorpos neutralizantes antivírus rábico, em amostra da população de Augusto Correa, vacinada com vacina de cultivo celular (Verorab®) por ocasião do surto. Foram coletadas e analisadas 505 amostras de moradores de quatro comunidades de Augusto Correa (Arai, Porto do Campo, Cachoeira e Nova Olinda) em junho de 2007. Os níveis de anticorpos neutralizantes foram dosados através da técnica de Favoretto e relacionados com dados demográficos, com história prévia de malária, novas agressões por animais e com o esquema vacinal recebido. Após dois anos da campanha vacinal, os resultados revelaram a persistência de anticorpos neutralizantes em níveis adequados em 90.5% da população vacinada, em esquemas de pré e pós-exposição, mesmo naqueles que receberam tratamento incompleto. Os níveis de anticorpos neutralizantes não foram reduzidos em função das variáveis analisadas.

Prevalence and geographical distribution of Toxoplasma gondii in dogs in the urban area of Botucatu, SP, Brazil

Toxoplasmosis is a worldwide zoonosis caused by Toxoplasma gondii, and can infect a wide variety of animals including humans. Domestic animals can be an important sentinel population for infection in the community. Occurrence of T. gondii infection was assessed in dogs in the urban area of Botucatu city, SP, Brazil. In the sample, 10% rate for error estimate, 95% confidence interval, and 5% significance level were established. Serum samples were collected from dogs during a rabies vaccination campaign, and were processed using modified agglutination test (MAT). Blood samples were collected from 670 dogs, with homogeneous distribution in five regions in the urban area, representing 3.74% of 17,910 animals vaccinated. In this sample, 17.3% (116/670) dogs (68 – 58.6% female and 48 – 41.4% male) were positive for T. gondii infection (p<0.03). Regarding age of the infected dogs 4.6% (4/88) were younger and 95.4% (84/88) were older than one year (p<0.01); the age of 28 positive animals were undetermined. The serum titers of anti-T. gondii antibodies were: 16 (69.8%; 81/116), 64 (13.8%; 16/116), 256 (15.5%; 18/116), and 1024 (0.9%; 1/116). Prevalence was distributed among the North 14.2% (19/134), South 18.0% (31/172), East 15.7% (19/121), West 21.6% (27/125), and Central 16.9% (20/118) regions of the municipality (p=0.5). In all these regions, females and dogs aged more than one year showed a higher occurrence of T. gondii infection (p<0.05).

A Field-Test of Rhodamine B as a Biomarker in Raccoons

Rhodamine B is a dye that when ingested results in fluorescent bands in growing hair and whiskers of many mammals. Previous research at Wildlife Services’ (WS) National Wildlife Research Center (NWRC) found that rhodamine B is a successful biomarker in raccoon whiskers and that raccoons do not have a taste aversion to the dye when it comprises ≤ 3% of a bait. Our study assessed the ease of bait distribution, whisker collection, and evaluation of the biomarker for potential use in the Oral Rabies Vaccination (ORV) program administered by the WS National Rabies Management Program (NRMP). In collaboration with WS operations personnel from Ohio and Michigan, 750 fishmeal polymer baits each containing 150 mg of rhodamine B were hand distributed at NASA’s Plum Brook Station, Sandusky, Ohio in the summer of 2008. Four weeks after baits were distributed whiskers from 162 raccoons were collected. Wildlife Services biologists and technicians evaluated the whiskers for fluorescence using a handheld UV magnifying lamp. Biologists then sent the whiskers to the NWRC, Ft. Collins, Colorado for confirmation of fluorescence under a UV microscope. Results suggest a high level of agreement between the two methods of evaluation. Surveys completed by biologists confirmed that the ease of use, less invasive sampling techniques and promptness of results obtained through the use of rhodamine B are advantageous to the tetracycline biomarker presently used by the ORV program. All participants recommended further evaluation of rhodamine B for its inclusion in future efforts requiring biomarker evaluation.

Effect of vaccination and the immunomodulators bacillus of Calmette-Guerin, Avridine and Propionibacterium acnes on rabies in mice

Responses: of vaccination and treatment to immunomodulators against rabies in mice were evaluated through macrophage inhibition factor (MIF), intra-pad inoculation (IPI) and serum neutralization (SN) tests and by the detection of gamma-interferon (IFN-gamma). Onco-BCG, Avridine and Propionibacterium acnes were administered to groups of mice. Higher survival rates were found in animals treated with P. acnes. Lower levels of IFN-gamma were observed in the groups of infected and vaccinated mice. The IPI was not effective on detecting the response of delayed-type hypersensitivity. Vaccine induced in the infected animals a more intense response to MIF reaction. (C) 1998 Elsevier B.V. Ltd. All rights reserved.

Rabies infection and specific effect of vaccination in mice selected for high and low immunobiological parameters

Innate and acquired resistance to rabies infection was investigated in mice genetically selected for high (H) or low (L) antibody responsiveness from selections I, III and IV and in mice selected for maximal (AIRmax) or minimal (AIRmin) acute inflammatory reaction. These mouse lines were infected intramuscularly with different virus dilutions and the LD50 was determined. The HIII and HIV mouse lines were more susceptible than the LIII and LIV lines and the HI line showed a discrete but higher resistance than the LI line. Analysis of the interline (H x L) F1 hybrids from selections III and IV indicated different dominance effects on the resistant and susceptible phenotypes when the route of vaccination was changed. No differences were observed between the AIRmax and AIRmin mice, suggesting that inflammation plays a minor role in the resistance to rabies virus. The comparison of LD50 in mice vaccinated by distinct routes showed that the highest interline difference occurred after intramuscular vaccination (250-fold between H and L and 800-fold between F1 and L). These results indicate that different mechanisms may participate in acquired antirabies resistance

Antibody and cytokine serum levels in patients subjected to anti-rabies prophylaxis with serum-vaccination

Rabies is considered a fatal disease once clinical symptoms have developed. The aim of this study was to evaluate epidemiological aspects and immune response in patients attacked by domestic and wild animals and subjected to post-exposure rabies treatment with equine serum and associated vaccine. Thirty-three patients were evaluated; they were between 13 and 65 years old, 75.8% were male and 24.2% female, and from the Botucatu neighborhood. Twenty healthy control individuals with the same age range were also studied. Specific antibodies to equine immunoglobulins and IFN-γ, IL-2, IL-4, and IL-10 production were evaluated by ELISA. IgM, IgE, IgG and subclasses, and rabies virus antibodies serum levels were determined by nephelometry and seroneutralization methods, respectively. No anaphylactic or serum sickness allergic reactions were observed in patients after treatment. Anti-equine IgG levels were significantly higher than those of IgM after 14 and 28 days of treatment. Protective antibodies to rabies virus > 0.5 UI/ml were detected in 84.6% and 75% of patients at days 14 and 28, respectively. IFN-γ, IL-2 and IL-10 levels in patients before and 48h after treatment were significantly higher than in controls suggesting that both Th1 and Th2 cells were activated in the patients. Serum IgM levels were higher at day 14, and IgG 2 and IgE levels were higher at day 28 of treatment. These results suggest that post-exposure rabies treatment in humans induces significant alterations in patient immune response characterized by increased levels of cytokines, serum levels of specific rabies virus antibodies, and the equine serum components employed in the treatment.

Development of broad-spectrum human monoclonal antibodies for rabies post-exposure prophylaxis

Currently available rabies post-exposure prophylaxis (PEP) for use in humans includes equine or human rabies immunoglobulins (RIG). The replacement of RIG with an equally or more potent and safer product is strongly encouraged due to the high costs and limited availability of existing RIG. In this study, we identified two broadly neutralizing human monoclonal antibodies that represent a valid and affordable alternative to RIG in rabies PEP. Memory B cells from four selected vaccinated donors were immortalized and monoclonal antibodies were tested for neutralizing activity and epitope specificity. Two antibodies, identified as RVC20 and RVC58 (binding to antigenic site I and III, respectively), were selected for their potency and broad-spectrum reactivity. In vitro, RVC20 and RVC58 were able to neutralize all 35 rabies virus (RABV) and 25 non-RABV lyssaviruses. They showed higher potency and breath compared to antibodies under clinical development (namely CR57, CR4098, and RAB1) and commercially available human RIG. In vivo, the RVC20–RVC58 cocktail protected Syrian hamsters from a lethal RABV challenge and did not affect the endogenous hamster post-vaccination antibody response.

Evaluation of cytokines concentration and percentage of survival of rabies virus-infected mice submitted to anti-rabies Vero-cell propagated vaccine and P-acnes

Previously, survival of rabies infection was shown to correlate with low IL-6 serum concentration in mice subjected to post-exposure treatment with the Fuenzalida Palacios rabies vaccine in conjunction with the immunomodulator Propionibacterium acnes, previously Corynebacterium parvum. Considering the substitution of the Fuenzalida Palacios rabies vaccine by the Vero cell raised anti-rabies vaccine in almost all countries, the objective of this work was to evaluate the survival and cytokine serum concentration of rabies virus-infected mice treated with P. acnes in conjunction with or the anti-rabies-VERO vaccine. For this, Swiss mice were experimentally infected with street rabies virus and subjected to vaccine and/or P. acnes following infection. Animals were killed at different times and serum was collected to evaluate cytokines. The greatest survival was observed in animals given one or two does of P. acnes in the absence of vaccination. Animals given anti-rabies VERO vaccine alone or with three doses of P. acnes had the second highest survival rate. The group that had the highest percentage of mortality also had the highest IL-6 concentration on the 10th day, a time correlating with clinical symptoms of the animals. The results reinforce the inefficacy of anti-rabies vaccine in only one dose as a post-exposure treatment irrespective of the type of vaccine used, the immunomodulation activity of P. acnes in rabies post-exposure treatment and suggest a role for IL-6 in rabies virus pathogenesis. (c) 2006 Elsevier B.V. All rights reserved.

Increased interleukin-10 associated with low IL-6 concentration correlated with greater survival rates in mice infected by rabies virus vaccinated against it and immunomodulated with P-acnes

Macrophage activity, cytokines serum concentration, serum neutralizing antibodies and lethality by rabies were evaluated in swiss mice experimentally infected with street rabies virus and submitted or not to antirabies vaccination and immunomodulation with P. acnes. Animals were killed at different times and serum was collected in order to evaluate cytokines concentration; peritonial and splenic macrophages were collected for macrophage activity evaluation. Greater survival rates higher IL-10 and low IL-6 serum concentration were observed in vaccinated animals treated using P. acnes. (C) 2004 Elsevier Ltd. All rights reserved.

Distribution of rabies virus in infected mice, vaccinated and submitted to P-acnes as immunomodulator

The lethality and distribution of rabies virus were evaluated in swiss mice experimentally infected with street rabies virus, vaccinated and submitted to immunomodulation by P. acnes (formerly Corynebacterium parvum). The animals were sacrificed at different times,when the different tissues were collected and submitted to fluorescent antibody test (FAT) and mouse inoculation test (MIT). The group submitted to vaccination and P. acnes treatment presented a percentage of survival superior to that observed in infected mice only treated with P. acnes. Control infected animals had the lowest survival rates. The distribution of rabies virus in spleen of infected mice, vaccinated and submitted to P. acnes was superior to that verified in infected mice not treated with P. acnes. The increased survival correlated with the distribution of rabies virus in lymphoid tissues, could be interpreted as the consequence of P. acnes activity on macrophages. The results suggest the role of macrophages against rabies virus infection in mice and the importance of vaccination in the post expositive treatment of rabies. (C) 2002 Published by Elsevier B.V. Ltd.

Efficiency of Matricaria chamomilla CH12 and number of doses of rabies vaccine on the humoral immune response in cattle

This study evaluated the effect of Matricaria chamomilla and vaccination frequency on cattle immunization against rabies. Four groups (n = 15/group) were treated with or without Matricaria chamomilla CH12 and vaccinated with one or two doses of rabies vaccine (30 day interval). No effect of chamomile was found on cattle immunization against rabies; however, antibody titers were protective in cattle vaccinated twice, while 93.3% of cattle vaccinated only once had titers under 0.5 UI/ml after 60 days. In conclusion, the use of chamomile did not alter the Immoral immune response in cattle, and two vaccine doses are suggested for achieving protective antibody titers.

Comparison between the counter immunoelectrophoresis test and mouse neutralization test for the detection of antibodies against rabies virus in dog sera

The detection of rabies antibodies is extremely valuable for epidemiological studies, determination of immune status in man, animals, and for the diagnosis of the disease. Several serological procedures have been described for this purpose. The present study reports a comparison between counterimmunoelectrophoresis test (CIET) and mouse neutralization test (MNT) in the detection of antibodies against rabies virus from 212 serum samples of vaccinated dogs. The agreement between both techniques was 79.7% and a significative association was demonstrated. The correlation coefficients between MNT and the CIET titers was determined considering 88 samples showing positive results in both techniques [CIET = 2 and MNT = 5 (0.13 IU/ml)] and resulted r² = 0.7926 (p < 0.001). The performance of CIET system was technically simple, cheap and rapid, and thereby it could be useful for serological monitoring of dog vaccination campaigns as well as for individual analysis.