Preemptive transcranial direct current stimulation induces analgesia, prevents chronic inflammation and fibrosis, and promotes tissue repair in a rat model of postoperative pain.

This study evaluated the effects of previous exposure to Transcranial Direct Current Stimulation (tDCS) on nociceptive, neuroinflammatory, and neurochemical parameters, in rats subjected to an incisional pain model. Forty adult male Wistar rats (60 days old; weighing âˆ¼ 250 g) were divided into five groups: 1. control (C); 2. drugs (D); 3. surgery (S); 4. surgery + sham-tDCS (SsT) and 5. surgery + tDCS (ST). Bimodal tDCS (0.5 mA) was applied for 20 min/day/8 days before the incisional model. Mechanical allodynia (von Frey) was evaluated at different time points after surgery. Cytokines and BDNF levels were evaluated in the cerebral cortex, hippocampus, brainstem, and spinal cord. Histology and activity of myeloperoxidase (MPO) and N-acetyl-ß-D-glucosaminidase (NAGase) were evaluated in the surgical lesion sites in the right hind paw. The results demonstrate that the surgery procedure increased BDNF and IL-6 levels in the spinal cord levels in the hippocampus, and decreased IL-1ß and IL-6 levels in the cerebral cortex, IL-6 levels in the hippocampus, and IL-10 levels in the brainstem and hippocampus. In addition, preemptive tDCS was effective in controlling postoperative pain, increasing BDNF, IL-6, and IL-10 levels in the spinal cord and brainstem, increasing IL-1ß in the spinal cord, and decreasing IL-6 levels in the cerebral cortex and hippocampus, IL-1ß and IL-10 levels in the hippocampus. Preemptive tDCS also contributes to tissue repair, preventing chronic inflammation, and consequent fibrosis. Thus, these findings imply that preemptive methods for postoperative pain management should be considered an interesting pain management strategy, and may contribute to the development of clinical applications for tDCS in surgical situations.

Repetitive Transcranial Magnetic Stimulation (rTMS) Reverses the Long-term Memory Impairment and the Decrease of Hippocampal Interleukin-10 Levels, both Induced by Neuropathic Pain in Rats.

Neuropathic pain (NP) is characterized by the presence of spontaneous pain, allodynia and hyperalgesia. Repetitive transcranial magnetic stimulation (rTMS) is one of neuromodulatory techniques that induces satisfactory NP relief, including that from refractory pain patients. The objective of this study was to evaluate rTMS treatment over long term memory (LTM) and hippocampal BDNF and IL-10 levels in rats submitted to a NP model. A total of 81 adult (60-days old) male Wistar rats were randomly allocated to one of the following 9 experimental groups: control, control + sham rTMS, control + rTMS, sham neuropathic pain, sham neuropathic pain + sham rTMS, sham neuropathic pain + rTMS, neuropathic pain (NP), neuropathic pain + sham rTMS and neuropathic pain + rTMS. Fourteen days after the surgery for chronic constriction injury (CCI) of the sciatic nerve, NP establishment was accomplished. Then, rats were treated with daily 5-minute sessions of rTMS for eight consecutive days. LTM was assessed by the object recognition test (ORT) twenty-four hours after the end of rTMS treatment. Biochemical assays (BDNF and IL-10 levels) were performed in hippocampus tissue homogenates. rTMS treatment reversed the reduction of the discrimination index in the ORT and the hippocampal IL-10 levels in NP rats. This result shows that rTMS reverses the impairment LTM and the increase in the hippocampal IL-10 levels, both induced by NP. Moreover, it appears to be a safe non-pharmacological therapeutic tool since it did not alter LTM and neurochemical parameters in naive animals.

Bimodal transcranial direct current stimulation reduces alcohol consumption and induces long-term neurochemical changes in rats with neuropathic pain.

This study aimed to evaluate the effects of repeated bimodal transcranial direct current stimulation (tDCS) on alcohol consumption and immunohistological and neurochemical parameters in nerve-injured rats. Forty-eight adult male Wistar rats were distributed into six groups: control, neuropathic pain (NP) + sham-tDCS, NP + alcohol + sham-tDCS, alcohol + sham-tDCS, alcohol + tDCS, and NP + alcohol + tDCS. NP is induced by chronic sciatic nerve constriction (CCI). The rats were exposed to a 10% alcohol solution by voluntary consumption for 14 days. From the 16th day after surgery, bimodal tDCS was applied for 20 min/day for 8 days. Brain structures were collected to evaluate the number of neuropeptide Y (NPY)-positive neurons, neurites, and argyrophilic grains by immunohistochemistry, and brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), interleukin (IL)-6, and IL-10 by ELISA. Nerve-injured rats showed a progressive increase in alcohol consumption compared to the non-injured rats. In addition, there was a reduction in voluntary alcohol consumption over time induced by tDCS. Alcohol exposure, chronic pain, and tDCS treatment modulated the central NPY immunoreactivity. tDCS increased the cerebellar levels of IL-6 and IL-10, and CCI and/or tDCS reduced striatal BDNF levels. The current data suggest that tDCS could be a promising non-pharmacological adjuvant to treat patients with chronic pain who use alcohol to relieve their symptoms.

rTMS induces analgesia and modulates neuroinflammation and neuroplasticity in neuropathic pain model rats.

Neuropathic pain (NP) is related to the presence of hyperalgesia, allodynia, and spontaneous pain, affecting 7%-10% of the general population. Repetitive transcranial magnetic stimulation (rTMS) is applied for NP relief, especially in patients with refractory pain. As NP response to existing treatments is often insufficient, we aimed to evaluate rTMS treatment on the nociceptive response of rats submitted to an NP model and its effect on pro-and anti-neuroinflammatory cytokine and neurotrophin levels. A total of 106 adult male Wistar rats (60 days old) were divided into nine experimental groups: control, control + sham rTMS, control + rTMS, sham NP, sham neuropathic pain + sham rTMS, sham neuropathic pain + rTMS, NP, neuropathic pain + sham rTMS, and neuropathic pain + rTMS. NP establishment was achieved 14 days after the surgery to establish chronic constriction injury (CCI) of the sciatic nerve. Rats were treated with 5 min daily sessions of rTMS for eight consecutive days. Nociceptive behavior was assessed using von Frey and hot plate tests at baseline, after NP establishment, and post-treatment. Biochemical assays to assess the levels of brain-derived neurotrophic factor (BDNF), tumor necrosis factor-alpha (TNF-α), and interleukin (IL)-10, were performed in the prefrontal cortex (PFC) and spinal cord tissue homogenates. rTMS treatment promoted a partial reversal of mechanical allodynia and total reversal of thermal hyperalgesia induced by CCI. Moreover, rTMS increased the levels of BDNF, TNF-α, and IL-10 in the PFC. rTMS may be a promising tool for the treatment of NP. The alterations induced by rTMS on neurochemical parameters may have contributed to the analgesic effect presented.

Intra- and inter-rater reliability in the assessment and classification of the longitudinal plantar arch of children 6 to 10 years of age

Abstract Aims: This study aimed to analyze the intra- and inter-rater reliability in the assessment and classification of the longitudinal plantar arch of children from 6 to 10 years old in the eyes-open (EO) testing condition. Methods: A total of two-hundred and seventy-eight Brazilian children (556 feet), boys and girls, from 6 to 10 years of age participated in the study. The children's feet were examined on a baropodometric platform, and the Staheli index was used for calculating the plantar arch index. Footprint analyses were performed at two different times, with an interval of 7 to 10 days, by three physical therapists in a single testing condition, resulting in 3,336 footprints. To determine the reliability of the continuous measurements, the Intraclass Correlation Coefficients (ICC) with 95% confidence intervals (CIs), Standard error of the mean (SEM), absolute value and percentage, and the Minimum Detectable Change (MDC) were calculated. To determine the reliability of the longitudinal arch classification, inter-rater reliability was evaluated by Weighted Fleiss Kappa Coefficient and the test-retest reliability was estimated by Weighted Cohen Kappa Coefficient. Results: Regarding inter-rater reliability, we observed values of ICC ranging from 0.79 to 0.96; thus, the results were classified as substantial to excellent reliability), being the lowest ICC values occur for line B, mainly in the first assessments. SEM ranges from 0.08 to 0.21 (percentage: 3.74 to 28.7), being the best, the lowest SEM values occur for Plantar Arch Index assessments and the MDC varies between 0.22 and 0.59. Regarding intra-rater reliability, the results indicated excellent reliability: values of ICC range from 0.92 to 0.99, being the lowest ICC values also occurs for line B analysis. SEM ranges from 0.03 to 0.20 (percentage: 2.32 to 26.6), being the lowest SEM values occur for Plantar Arch Index assessments and MDC varies between 0.09 and 0.54. Analyzing the inter-rater reliability for the longitudinal arch classifications, we observed values of Weighted Fleiss Kappa Coefficient ranging from 0.83 to 0.87, expressing almost perfect agreement among the raters before and after evaluations. The test-retest reliability of the longitudinal arch classification resulted in values of Weighted Cohen Kappa Coefficient ranging from 0.80 to 0.996, expressing substantial to almost perfect agreement intra-rater. Conclusion: The study showed high reliability in the clinical assessment of the longitudinal plantar arch index of children from 6 to 10 years of age indicating that the Staheli method is applicable to pressure platform assessments with intra- and inter-rate reliability.

Nicotinamide riboside reduces cardiometabolic risk factors and modulates cardiac oxidative stress in obese Wistar rats under caloric restriction.

AIMS: NAD-based therapeutic strategies are encouraged against obesity and heart disease. Our study, therefore, aimed to investigate the effects of nicotinamide riboside (NR), isolated or combined with caloric restriction (CR), both approaches well-known for stimulating NAD levels, on adiposity parameters, cardiometabolic factors and cardiac oxidative stress in rats submitted to cafeteria diet (CAF). MAIN METHODS: After 42 days of CAF-induced obesity (hypercaloric and ultra-processed foods common to humans), we examined the effects of oral administration of NR (400 mg/kg for 28 days), combined or not with CR (-62% kcal, for 28 days), on anthropometric, metabolic, tissue, and cardiac oxidative stress parameters in obese male Wistar rats. KEY FINDINGS: In obese rats, treatment with NR alone mitigated final body weight gain, reduced adiposity (visceral and subcutaneous), improved insulin resistance, and decreased TG/HDL ratio and heart size. In cardiac OS, treatment with NR increased the antioxidant capacity via glutathione peroxidase and catalase enzymes (in rats under CR) as well as reduced the pro-oxidant complex NADPH oxidase (in obese and lean rats). Hyperglycemia, hypertriglyceridemia and elevated levels of TBARS in the heart were state-dependent adverse effects, induced by treatment with NR. SIGNIFICANCE: This is the first study to report effects of nicotinamide riboside on cardiac oxidative stress in an obesity model. Nicotinamide riboside, a natural dietary compound, presented antiobesity effects and cardiometabolic benefits, in addition to positively modulating oxidative stress in the heart, in a state-dependent manner.

Neonatal morphine exposure and maternal deprivation alter nociceptive response and central biomarkers' levels throughout the life of rats.

In the present study, we investigated the effect of repeated neonatal morphine exposure and/or maternal deprivation(MD) on the nociceptive response and central biomarkers' BDNF, IL-1ß, and IL-4 levels at postnatal days 16(PND16), 30(PND30), and 60(PND60). At birth, the litters were standardized to contain 8 pups/dam (n = 58). From PND1 to PND10, the pups of the deprived groups were separated daily from their mothers for 3 h and divided into 5 groups: control(C), saline(S), morphine(M), deprived-saline(DS), and deprived-morphine(DM). The pups received subcutaneous injections of saline/morphine (5 µg) in the mid-scapular area between PND8 and PND14. Nociceptive responses were assessed by hot plate(HP) and tail-flick(TFL) tests and biomarker levels by ELISA. Thermal hyperalgesia(HP) was found in all assessments for the M, DS, and DM groups, and a decrease in nociceptive threshold(TFL) was found in the DS group at PND16; M and DM groups at PND30; and M, DS, and DM groups at PND60. There were interactions between treatment/deprivation/timepoint in all central biomarkers' levels. The current study indicates that neonatal exposure to morphine and MD, which occurs in the pediatric ICU, can alter the nociceptive and neuroinflammatory responses.

Association between diabetes and vestibular dysfunction: an integrative review / Associação entre diabetes e disfunções do sistema vestibular: revisão integrativa

ABSTRACT Purpose: to identify, in the literature, the factors associated with the development of vestibular dysfunctions in individuals with type 2 diabetes mellitus (DM2). Methods: an integrative review of the literature, whose survey was conducted in the databases ISI, SciELO, LILACS and PubMed, using the following descriptors: "type 2 diabetes mellitus", "vertigo", "dizziness", and "vestibular diseases". Articles published in the last 10 years that answered the research question ("What factors are associated with the development of vestibular disorders in individuals with DM2?") were included in the study. Results: the search returned 426 articles, 10 of which met the eligibility criteria. Most of the participants of the selected studies who had vestibular dysfunctions were women over 40 years old and had more than one comorbidity related to DM2, the main one being the systemic arterial hypertension (SAH). According to the literature, the physiology of the inner ear allows small glucose alterations to influence its normal functioning, which makes diabetic individuals more susceptible to developing vestibular dysfunctions. Conclusion: according to this study, DM2 can trigger or contribute to the manifestation of vestibular dysfunction, whose main associated factors are advanced age, female gender, and various comorbidities, as dyslipidemia, SAH and metabolic syndrome.

RESUMO Objetivo: identificar na literatura os fatores que estão associados ao desenvolvimento de disfunções vestibulares em indivíduos com diabetes mellitus tipo 2 (DM2). Métodos: trata-se de uma revisão integrativa da literatura, cuja busca foi realizada nas bases de dados ISI, Scielo, LILACS e PubMed, sendo utilizados os descritores: "type 2 diabetes mellitus", "vertigo", "dizziness" e "vestibular diseases". Foram incluídos artigos publicados nos últimos dez anos e que respondessem a seguinte questão: quais fatores estão associados ao desenvolvimento de disfunções vestibulares em indivíduos com DM2? Resultados: a busca resultou em 426 artigos, sendo que 10 atenderam aos critérios de elegibilidade. A maioria dos participantes dos estudos selecionados que apresentaram disfunções vestibulares eram do sexo feminino, idade superior a 40 anos e possuíam mais de uma comorbidade associada a DM2, sendo a principal, a hipertensão arterial sistêmica (HAS). De acordo com a literatura, a fisiologia da orelha interna permite que pequenas alterações de glicose influenciem seu funcionamento normal, o que torna os indivíduos diabéticos mais suscetíveis a desenvolverem disfunções vestibulares. Conclusão: o presente estudo constatou que a DM2 pode desencadear ou contribuir para a manifestação da disfunção vestibular, tendo como principais fatores associados a idade avançada, o sexo feminino e comorbidades diversas, tais como dislipidemia, HAS e síndrome metabólica.

Surveillance of Giardia and Cryptosporidium in sewage from an urban area in Brazil.

Cryptosporidium and Giardia are protozoan parasites that cause diarrhea in humans and animals. Molecular characterization of these pathogens in sewage may provide insight on their occurrence and prevalence in Brazil. This study aimed to investigate the presence of Giardia and Cryptosporidium in raw and treated sewage from Londrina, Paraná, Brazil. Samples were collected every two weeks during a year. Samples were concentrated, then DNA was extracted and subjected to a nested PCR targeting the Giardia 18S rRNA gene and the Cryptosporidium 18S rRNA gene. Species of Cryptosporidium were characterized by restriction fragment length polymorphism (RFLP). All raw sewage and 76% of the treated sewage were positive for Giardia; 84% of raw sewage samples and 8% of treated sewage were positive for Cryptosporidium. C. muris, C. hominis, C. baileyi, C. parvum and C. suis were detected in 100%, 19%, 9%, 9% and 4% of raw sewage, respectively. C. muris was the only species found in treated sewage. Multiple species of Cryptosporidium were present in 19.04% of the raw sewage. Treated sewage water can pose a threat to human health. The speciation of Cryptosporidium revealed the presence of non-common zoonotic species as C. suis and C. muris.

Surveillance of Giardia and Cryptosporidium in sewage from an urban area in Brazil / Monitoramento de Giardia e Cryptosporidium em esgoto de uma área urbana no Brasil

Abstract Cryptosporidium and Giardia are protozoan parasites that cause diarrhea in humans and animals. Molecular characterization of these pathogens in sewage may provide insight on their occurrence and prevalence in Brazil. This study aimed to investigate the presence of Giardia and Cryptosporidium in raw and treated sewage from Londrina, Paraná, Brazil. Samples were collected every two weeks during a year. Samples were concentrated, then DNA was extracted and subjected to a nested PCR targeting the Giardia 18S rRNA gene and the Cryptosporidium 18S rRNA gene. Species of Cryptosporidium were characterized by restriction fragment length polymorphism (RFLP). All raw sewage and 76% of the treated sewage were positive for Giardia; 84% of raw sewage samples and 8% of treated sewage were positive for Cryptosporidium. C. muris, C. hominis, C. baileyi, C. parvum and C. suis were detected in 100%, 19%, 9%, 9% and 4% of raw sewage, respectively. C. muris was the only species found in treated sewage. Multiple species of Cryptosporidium were present in 19.04% of the raw sewage. Treated sewage water can pose a threat to human health. The speciation of Cryptosporidium revealed the presence of non-common zoonotic species as C. suis and C. muris.

Resumo Cryptosporidium e Giardia são protozoários causadores de diarreia em animais e humanos. A caracterização molecular destes protozoários em esgoto pode prover dados ainda desconhecidos da ocorrência de espécies. O objetivo do presente estudo foi monitorar a ocorrência de Giardia e espécies de Cryptosporidium em esgoto bruto e tratado em uma estação de tratamento de esgoto (ETE) de Londrina, Paraná. Amostras de esgoto bruto e tratado foram coletadas no período de um ano, com periodicidade quinzenal. A ocorrência destes protozoários foi caracterizada por meio de concentração das amostras e posterior extração de DNA seguida de nested-PCR para amplificação de fragmentos dos genes 18S rRNA de Giardia e 18S rRNA de Cryptosporidium. A caracterização das espécies de Cryptosporidium foi realizada por meio de análise por polimorfismo de comprimento do fragmento de restrição (RFLP) dos produtos obtidos. Foram coletadas no total 25 amostras de cada, esgoto bruto e esgoto tratado. Para Giardia, todas as amostras de esgoto bruto e 76% das de esgoto tratado foram positivas. Cryptosporidium esteve presente em 84% das amostras de esgoto bruto e em 8% do tratado. No esgoto tratado foi encontrado apenas C. muris, já nas amostras de esgoto bruto foram encontradas cinco espécies: C. muris, C. hominis, C. baileyi, C. suis e C. parvum em 100%, 19%, 9%, 9% e 4%, respectivamente. A presença de espécies mistas foi observada em 19,04% das amostras. A presença de Giardia e Cryptosporidium em esgoto tratado pode pôr em risco a saúde humana. A discriminação de espécies de Cryptosporidium revelou a presença de espécies zoonóticas incomuns como C. suis e C. muris.

Cryptosporidium spp. and Giardia spp. in feces and water and the associated exposure factors on dairy farms.

The aims of this study were to verify the prevalence of Cryptosporidium spp. and Giardia spp. in animal feces and drinking water on dairy farms and to identify a possible relation between the exposure factors and the presence of these parasites. Fecal samples from cattle and humans and water samples were collected on dairy farms in Paraná, Brazil. Analysis of (oo)cysts in the feces was performed by the modified Ziehl-Neelsen staining and centrifugal flotation in zinc sulfate. Test-positive samples were subjected to nested PCR amplification of the 18SSU ribosomal RNA gene for identification of Cryptosporidium and Giardia and of the gp60 gene for subtyping of Cryptosporidium. Microbiological analysis of water was carried out by the multiple-tube method and by means of a chromogenic substrate, and parasitological analysis was performed on 31 samples by direct immunofluorescence and nested PCR of the genes mentioned above. Identification of the species of Cryptosporidium was performed by sequencing and PCR with analysis of restriction fragment length polymorphisms. The prevalence of Giardia and Cryptosporidium was higher in calves than in adults. Among the samples of cattle feces, Cryptosporidium parvum was identified in 41 (64%), C. ryanae in eight (12.5%), C. bovis in four (6.3%), C. andersoni in five (7.8%), and a mixed infection in 20 samples (31.3%). These parasites were not identified in the samples of human feces. Thermotolerant coliform bacteria were identified in 25 samples of water (45.5%). Giardia duodenalis and C. parvum were identified in three water samples. The gp60 gene analysis of C. parvum isolates revealed the presence of two strains (IIaA20G1R1 and IIaA17G2R2) in the fecal samples and one (IIaA17G2R1) in the water samples. The presence of coliforms was associated with the water source, structure and degradation of springs, rain, and turbidity. The prevalence of protozoa was higher in calves up to six months of age. C. parvum and G. duodenalis were identified in the water of dairy farms, as were thermotolerant coliforms; these findings point to the need for guidance on handling of animals, preservation of water sources, and water treatment.

Serosurvey of Borrelia in dogs, horses, and humans exposed to ticks in a rural settlement of southern Brazil.

The aims of the present study were to serosurvey dogs, horses, and humans highly exposed to tick bites for anti-Borrelia burgdorferi s.l. antibodies, identify tick species present, and determine risk factors associated with seropositivity in a rural settlement of Paraná State, southern Brazil. Eighty-seven residents were sampled, along with their 83 dogs and 18 horses, and individual questionnaires were administered. Immunofluorescence antibody test (IFAT) was performed on serum samples and positive samples were subjected to western blot (WB) analysis. Anti-B. burgdorferi antibodies were found in 4/87 (4.6%) humans, 26/83 (31.3%) dogs, and 7/18 (38.9%) horses by IFAT, with 4/4 humans also positive by WB. Ticks identified were mostly from dogs and included 45/67 Rhipicephalus sanguineus, 21/67 Amblyomma ovale, and 1/67 A. cajennense sensu lato. All (34/34) horse ticks were identified as A. cajennense s.l.. No significant association was found when age, gender, or presence of ticks was correlated to seropositivity to Borrelia sp. In conclusion, although anti-Borrelia antibodies have been found in dogs, horses and their owners from the rural settlement, the lack of isolation, molecular characterization, absence of competent vectors and the low specificity of the commercial WB kit used herein may have impaired risk factor analysis.

Serosurvey of Borrelia in dogs, horses, and humans exposed to ticks in a rural settlement of southern Brazil / Soroprevalência e fatores associados a Borrelia em cães, equinos e humanos expostos a carrapatos em um assentamento rural do sul do Brasil

Abstract The aims of the present study were to serosurvey dogs, horses, and humans highly exposed to tick bites for anti-Borrelia burgdorferi s.l. antibodies, identify tick species present, and determine risk factors associated with seropositivity in a rural settlement of Paraná State, southern Brazil. Eighty-seven residents were sampled, along with their 83 dogs and 18 horses, and individual questionnaires were administered. Immunofluorescence antibody test (IFAT) was performed on serum samples and positive samples were subjected to western blot (WB) analysis. Anti-B. burgdorferi antibodies were found in 4/87 (4.6%) humans, 26/83 (31.3%) dogs, and 7/18 (38.9%) horses by IFAT, with 4/4 humans also positive by WB. Ticks identified were mostly from dogs and included 45/67 Rhipicephalus sanguineus, 21/67 Amblyomma ovale, and 1/67 A. cajennense sensu lato. All (34/34) horse ticks were identified as A. cajennense s.l.. No significant association was found when age, gender, or presence of ticks was correlated to seropositivity to Borrelia sp. In conclusion, although anti-Borrelia antibodies have been found in dogs, horses and their owners from the rural settlement, the lack of isolation, molecular characterization, absence of competent vectors and the low specificity of the commercial WB kit used herein may have impaired risk factor analysis.

Resumo Os objetivos do presente estudo foram realizar um levantamento sorológico de cães, cavalos e humanos altamente expostos a picadas de carrapatos para anticorpos anti-B. burgdorferi s.l., identificar as espécies de carrapatos presentes, e determinar os fatores de risco associados a soropositividade em um assentamento rural do Estado do Paraná, sul do Brasil. Oitenta e sete residentes foram amostrados junto com seus respectivos 83 cães e 118 cavalos e questionários individuais foram aplicados. O teste de imunofluorescência indireta (IFI) foi realizado nas amostras sorológicas e as positivas foram submetidas a análise por western blot (WB). Anticorpos anti-B. burgdorferi foram detectados em 4/87 (4,6%) humanos, 26/83 (31,3%) cães e 7/18 (38,9%) cavalos pela IFI, com 4/4 humanos também positivos pelo WB. Os carrapatos identificados foram em sua maioria de cães e incluíram 45/67 Rhipicephalus sanguineus, 21/67 Amblyomma ovale e 1/67 A. cajennense sensu lato. Todos (34/34) carrapatos dos cavalos foram identificados como A. cajennense s.l.. Não foram observadas diferenças estatísticas entre idade, sexo ou presença de carrapatos e soropositividade para Borrelia sp. Em conclusão, embora anticorpos anti-Borrelia tenham sido encontrados em cães, equinos e seus proprietários do assentamento rural, a ausência de isolamento, caracterização molecular, ausência de vetores competentes e baixa especificidade do kit comercial de WB utilizado podem ter limitado a análise de fatores de risco.

ISOLATION AND GENOTYPING OF Toxoplasma gondii IN SERONEGATIVE URBAN RATS AND PRESENCE OF ANTIBODIES IN COMMUNICATING DOGS IN BRAZIL.

The role of rodents in the epidemiology of toxoplasmosis was investigated in Londrina, Paraná State, Brazil. One hundred and eighty-one Rattus rattus and one Mus musculus were caught in 37 places. Blood and tissues were collected and submitted to the indirect fluorescence antibody test (IFAT) and the bioassay. Serum samples from 61 contacting dogs were also collected. Sixteen rats (8.8%) were positive for Toxoplasma gondii, but just two of them were positive by serology and bioassay test. Antibodies were found in nine (4.9%) rats. Tissues of nine rats bioassayed were positive and four isolates were obtained. Restriction fragment length polymorphism (RFLP) analysis was performed using 12 markers (SAG1, SAG2, SAG2-alt, C22-8, C29-2, L358, PK1, BTUB, GRA6, SAG3, Apico, CS3). Genotyping revealed that the four strains isolated from this study have been isolated before in cats and chickens from Brazil. None of the isolates was identified like clonal archetypal T-types I, II, and III. The rats presented lower serologic Toxoplasma gondii prevalence (8.8%) compared to contacting dogs (70.5%).

Serosurvey of tick-borne pathogens in dogs from urban and rural areas from Parana State, Brazil.

Considering the zoonotic potential of tick-borne disease (TBD) agents and the fact that dogs may act as sentinels for human infection, the aim of the present study was to determine the seroprevalence of TBD agents and risk factors for exposure in two different canine populations from Parana State, Southern Brazil. A total of 138 dog serum samples from urban (UA) (n=68) and rural (RA) (n=70) areas were tested with commercial ELISA rapid test for Anaplasma phagocytophilum, Ehrlichia canis and Borrelia burgdorferi antibodies and indirect immunofluorescence assay (IFAT) for Babesia vogeli. An overall of 92∕138 (66.7%) dogs, being 62∕68 (91.2%) from UA and 30∕70 (42.9%) from RA, were seropositive for at least one TBD agent. From the total number of dogs, sixty-two were positive for E. canis (44.9%), 19 (13.8%) for A. phagocytophilum, and 64 (46.4%) for B. vogeli. Anti-B. burgdorferi antibodies were not detected. Dogs from UA showed a higher percentage of tick infestation (p = 0.0135) and were highly associated with seropositivity to E. canis (p = 0.000005), A. phagocytophilum (p = 0.0001), and B. vogeli (p = 0.0012). In summary, the findings indicate that dogs from urban areas present higher potential risk exposure to TBD pathogens than those from rural areas.

Serosurvey of tick-borne pathogens in dogs from urban and rural areas from Parana State, Brazil / Avaliação sorológica de patógenos transmitidos por carrapatos em cães urbanos e rurais do estado do Paraná, Brasil

Considering the zoonotic potential of tick-borne disease (TBD) agents and the fact that dogs may act as sentinels for human infection, the aim of the present study was to determine the seroprevalence of TBD agents and risk factors for exposure in two different canine populations from Parana State, Southern Brazil. A total of 138 dog serum samples from urban (UA) (n=68) and rural (RA) (n=70) areas were tested with commercial ELISA rapid test forAnaplasma phagocytophilum, Ehrlichia canisand Borrelia burgdorferi antibodies and indirect immunofluorescence assay (IFAT) for Babesia vogeli. An overall of 92∕138 (66.7%) dogs, being 62∕68 (91.2%) from UA and 30∕70 (42.9%) from RA, were seropositive for at least one TBD agent. From the total number of dogs, sixty-two were positive for E. canis (44.9%), 19 (13.8%) for A. phagocytophilum, and 64 (46.4%) for B. vogeli. Anti-B. burgdorferi antibodies were not detected. Dogs from UA showed a higher percentage of tick infestation (p = 0.0135) and were highly associated with seropositivity to E. canis (p = 0.000005), A. phagocytophilum (p = 0.0001), and B. vogeli (p = 0.0012). In summary, the findings indicate that dogs from urban areas present higher potential risk exposure to TBD pathogens than those from rural areas.

Considerando o potencial zoonótico das doenças transmitidas por carrapatos (DTCs) e que os cães podem atuar como sentinelas para infecções em humanos, os objetivos deste estudo foram determinar a soroprevalência de agentes das DTCs e fatores de risco para a exposição em duas diferentes populações caninas do Estado do Paraná, região Sul do Brasil. Um total de 138 amostras de soro de cães de área urbana (AU) (n = 68) e rural (AR) (n = 70) foram testadas utilizando um teste de ELISA comercial rápido para detecção de anticorpos contra Anaplasma phagocytophilum, Ehrlichia canis e Borrelia burgdorferi e imunofluorescência indireta (IFI) para Babesia vogeli. Um total de 92∕138 (66,7%) cães, sendo 62∕68 (91,2%) da AU e 30∕70 (42,9%) da AR, foram soropositivos para pelo menos um agente. Do número total de amostras, sessenta e duas (44,9%) foram positivas para E. canis, 19 (13,8%) para A. phagocytophilum e 64 (46,4%) para B. canis vogeli. Anticorpos anti-B. burgdorferi não foram detectados. Os cães da AU apresentaram o maior percentual de infestação por carrapatos (p = 0,0135) e foram altamente associados com a positividade para E. canis (p = 0,000005), A. phagocytophilum (p = 0,0001) e B. vogeli (p = 0,0012). Em resumo, nossos achados indicam que cães de áreas urbanas têm um maior risco potencial de exposição a agentes patogênicos das DTCs comparados aos das áreas rurais.

Study of infection by Rickettsiae of the spotted fever group in humans and ticks in an urban park located in the City of Londrina, State of Paraná, Brazil.

INTRODUCTION: Spotted fevers are emerging zoonoses caused by Rickettsia species in the spotted fever group (SFG). Rickettsia rickettsii is the main etiologic agent of Brazilian spotted fever (BSF)and it is transmitted by Amblyomma spp. ticks. METHODS: The study aimed to investigate SFG rickettsiae in the Arthur Thomas Municipal Park in Londrina, PR, by collecting free-living ticks and ticks from capybaras and blood samples from personnel working in these areas. Samples from A. dubitatum and A. cajennense were submitted for PCR in pools to analyze the Rickettsia spp. gltA (citrate synthase gene). RESULTS: All the pools analyzed were negative. Human sera were tested by indirect immunofluorescence assay with R. rickettsii and R. parkeri as antigens.Among the 34 sera analyzed, seven (20.6%) were reactive for R. rickettsii: four of these had endpoint titers equal to 64, 2 titers were 128 and 1 titer was 256. None of the samples were reactive for R. parkeri. An epidemiological questionnaire was applied to the park staff, but no statistically significant associations were identified. CONCLUSIONS: The serological studies suggest the presence of Rickettsiae related to SFG that could be infecting the human population studied; however, analysis of the ticks collected was unable to determine which species maybe involved in transmission to humans.

Study of infection by Rickettsiae of the spotted fever group in humans and ticks in an urban park located in the City of Londrina, State of Paraná, Brazil / Estudo da infecção por Rickettsias do grupo da febre maculosa em humanos e carrapatos de um parque urbano na Cidade de Londrina, Estado do Paraná

INTRODUCTION: Spotted fevers are emerging zoonoses caused by Rickettsia species in the spotted fever group (SFG). Rickettsia rickettsii is the main etiologic agent of Brazilian spotted fever (BSF) and it is transmitted by Amblyomma spp. ticks. METHODS: The study aimed to investigate SFG rickettsiae in the Arthur Thomas Municipal Park in Londrina, PR, by collecting free-living ticks and ticks from capybaras and blood samples from personnel working in these areas. Samples from A. dubitatum and A. cajennense were submitted for PCR in pools to analyze the Rickettsia spp. gltA (citrate synthase gene). RESULTS: All the pools analyzed were negative. Human sera were tested by indirect immunofluorescence assay with R. rickettsii and R. parkeri as antigens. Among the 34 sera analyzed, seven (20.6 percent) were reactive for R. rickettsii: four of these had endpoint titers equal to 64, 2 titers were 128 and 1 titer was 256. None of the samples were reactive for R. parkeri. An epidemiological questionnaire was applied to the park staff, but no statistically significant associations were identified. CONCLUSIONS: The serological studies suggest the presence of Rickettsiae related to SFG that could be infecting the human population studied; however, analysis of the ticks collected was unable to determine which species may be involved in transmission to humans.

INTRODUÇÃO: A febre maculosa é uma zoonose emergente causada por espécies de Rickettsia do grupo febre maculosa (GFM). Rickettsia rickettsii é o principal agente etiológico da febre maculosa brasileira (FMB) e é transmitida por Amblyomma spp. MÉTODOS: Com o objetivo de obter informações sobre GFM Rickettsiae no Parque Municipal Arthur Thomas em Londrina, PR, carrapatos de vida livre e de capivaras foram coletados, assim como amostras de sangue das pessoas que trabalham no parque. A. dubitatum e A. cajennense foram submetidos à PCR em pools para analises de Rickettsia spp. gltA (citrate synthase gene). RESULTADOS: Todos os pools de carrapatos analizados foram negativos. Soros de humanos foram testados pela imunofluorescência indireta com antigenos de R. rickettsii e R. parkeri. Entre os 34 soros analisados, 7 (20,6 por cento) foram positivos para R. rickettsii. Destes, quatro apresentaram títulos iguais a 64, dois iguais a 128 e um, igual a 256, mas nenhum soro reagiu com R. parkeri. Não houve nenhuma associação, estatisticamente significante, entre as variáveis analisadas no questionário epidemiológico fornecido às pessoas que participaram da pesquisa. CONCLUSÕES: Os estudos sorológicos sugerem a presença de alguma Rickettsiae relacionada ao GFM que poderiam estar infectando a população humana estudada. Entretanto, as análises dos carrapatos foram inconclusivas para determinar qual espécie poderia estar envolvida na transmissão para os humanos.