Parasite load in the blood and skin of dogs naturally infected by Leishmania infantum is correlated with their capacity to infect sand fly vectors

AbstractThe sand fly Lutzomyia longipalpis is primarily responsible for the transmission of visceral leishmaniasis (VL) in the New World, and dogs are considered to be the main urban reservoir of this disease. In order to improve the efficacy of control measures, it is essential to assess the transmission capacity of Leishmania infantum to the sand fly vector by naturally infected dogs. The present study investigated the existence of correlations between canine clinical presentation and the intensity of parasite load in the blood, skin and spleen of naturally infected dogs. In addition, we also attempted to establish correlations between the intensity of parasite load in canine tissue and the parasite load detected in sandflies five days after feeding on naturally infected dogs. A total of 23 dogs were examined and classified according to clinical manifestation of canine VL. Blood samples, splenic aspirate and skin biopsies were collected and parasite DNA was quantified by qPCR. Canine capacity to infect Lu. longipalpis with parasites was evaluated by xenodiagnosis and parasite loads were measured five days after feeding. No significant differences were observed with respect to canine clinical manifestation and the parasite loads detected in the blood, skin and spleen samples obtained from naturally infected dogs. Regardless of clinical canine visceral leishmaniasis (CVL) presentation and the degree of parasite burden, almost half of the dogs successfully infected sandflies with parasites, albeit to a low number of sandflies with correspondingly low parasite loads. Parasite loads in both canine blood and skin were shown to be positively correlated with the canine infectiousness to the sand fly vector, and positive correlations were also observed with respect to these tissues and the sand fly infection rate, as well as the parasite load detected in sandflies following xenodiagnosis. In conclusion, this indicates that parasite loads in both blood and skin can function as potentially reliable markers of canine capacity to infect sand fly vector

Cholinergic Signaling Exerts Protective Effects in Models of Sympathetic Hyperactivity-Induced Cardiac Dysfunction

Cholinergic control of the heart is exerted by two distinct branches; the autonomic component represented by the parasympathetic nervous system, and the recently described non-neuronal cardiomyocyte cholinergic machinery. Previous evidence has shown that reduced cholinergic function leads to deleterious effects on the myocardium. Yet, whether conditions of increased cholinergic signaling can offset the pathological remodeling induced by sympathetic hyperactivity, and its consequences for these two cholinergic axes are unknown. Here, we investigated two models of sympathetic hyperactivity: i) the chronic beta-adrenergic receptor stimulation evoked by isoproterenol (ISO), and ii) the alpha(2A)/alpha(2C)-drenergic receptor knockout (KO) mice that lack pre-synaptic adrenergic receptors. In both models, cholinergic signaling was increased by administration of the cholinesterase inhibitor, pyridostigmine. First, we observed that isoproterenol produces an autonomic imbalance characterized by increased sympathetic and reduced parasympathetic tone. Under this condition transcripts for cholinergic proteins were upregulated in ventricular myocytes, indicating that non-neuronal cholinergic machinery is activated during adrenergic overdrive. Pyridostigmine treatment prevented the effects of ISO on autonomic function and on the ventricular cholinergic machinery, and inhibited cardiac remodeling. alpha(2A)/alpha(2C)-KO mice presented reduced ventricular contraction when compared to wild-type mice, and this dysfunction was also reversed by cholinesterase inhibition. Thus, the cardiac parasympathetic system and non-neuronal cardiomyocyte cholinergic machinery are modulated in opposite directions under conditions of increased sympathetic drive or ACh availability. Moreover, our data support the idea that pyridostigmine by restoring ACh availability is beneficial in heart disease

Succinate causes pathological cardiomyocyte hypertrophy through GPR91 activation

Background\ud Succinate is an intermediate of the citric acid cycle as well as an extracellular circulating molecule, whose receptor, G protein-coupled receptor-91 (GPR91), was recently identified and characterized in several tissues, including heart. Because some pathological conditions such as ischemia increase succinate blood levels, we investigated the role of this metabolite during a heart ischemic event, using human and rodent models.\ud \ud \ud Results\ud We found that succinate causes cardiac hypertrophy in a GPR91 dependent manner. GPR91 activation triggers the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), the expression of calcium/calmodulin dependent protein kinase IIδ (CaMKIIδ) and the translocation of histone deacetylase 5 (HDAC5) into the cytoplasm, which are hypertrophic-signaling events. Furthermore, we found that serum levels of succinate are increased in patients with cardiac hypertrophy associated with acute and chronic ischemic diseases.\ud \ud \ud Conclusions\ud These results show for the first time that succinate plays an important role in cardiomyocyte hypertrophy through GPR91 activation, and extend our understanding of how ischemia can induce hypertrophic cardiomyopathy.CAPESFAPEMIG (Pronex)INCT- Carbon NanotubesCNPqHHM

Eletrocardiografia computadorizada em cães: estudo comparativo

O método de eletrocardiografia computadorizada (ECG-C) vem sendo crescentemente difundido na medicina veterinária, havendo atualmente diversas marcas e modelos de eletrocardiógrafos disponíveis no mercado. Diante da possibilidade de diferenças na sensibilidade e na reprodutibilidade das medidas obtidas nos traçados, o presente estudo teve como objetivo comparar os parâmetros eletrocardiográficos de cães, obtidos por dois sistemas. Foram avaliados dois diferentes softwares computadorizados, o Wincardio Micromed® (WIN) e o modelo TEB ECGPC® (TEB). Quarenta e dois cães hígidos, de diferentes raças (Cocker Spaniel, Daschund, Labrador, Pinscher, Pit Bull Terrier Poodle, Schnauzer, Shit Tzu, Yorkshire e sem raça definida), machos e fêmeas e com idade entre 4 meses e 16 anos foram agrupados segundo o peso e examinados pelos dois sistemas. As medidas eletrocardiográficas dos diferentes traçados foram analisadas na derivação DII. Os resultados indicaram que o sistema TEB apresentou maior sensibilidade na obtenção das medidas de duração da onda P e do complexo QRS, enquanto o sistema WIN foi mais sensível para determinar as medidas de amplitude dos mesmos parâmetros. Os animais de maior porte (26-37kg) apresentaram maior variância nas medidas de duração e amplitude de onda P e duração do complexo QRS em comparação aos cães de médio (14-25kg) e pequeno (1-13kg) porte. O achado de diferenças entre os sistemas testados deve ser levado em consideração ao se empregar os diversos equipamentos para diagnóstico por meio de ECG-C na rotina clínica, de modo a evitarem-se divergências na interpretação dos exames entre diferentes prestadores de serviços veterinários

Eletrocardiografia computadorizada em cães: estudo comparativo

O método de eletrocardiografia computadorizada (ECG-C) vem sendo crescentemente difundido na medicina veterinária, havendo atualmente diversas marcas e modelos de eletrocardiógrafos disponíveis no mercado. Diante da possibilidade de diferenças na sensibilidade e na reprodutibilidade das medidas obtidas nos traçados, o presente estudo teve como objetivo comparar os parâmetros eletrocardiográficos de cães, obtidos por dois sistemas. Foram avaliados dois diferentes softwares computadorizados, o Wincardio Micromed® (WIN) e o modelo TEB ECGPC® (TEB). Quarenta e dois cães hígidos, de diferentes raças (Cocker Spaniel, Daschund, Labrador, Pinscher, Pit Bull Terrier Poodle, Schnauzer, Shit Tzu, Yorkshire e sem raça definida), machos e fêmeas e com idade entre 4 meses e 16 anos foram agrupados segundo o peso e examinados pelos dois sistemas. As medidas eletrocardiográficas dos diferentes traçados foram analisadas na derivação DII. Os resultados indicaram que o sistema TEB apresentou maior sensibilidade na obtenção das medidas de duração da onda P e do complexo QRS, enquanto o sistema WIN foi mais sensível para determinar as medidas de amplitude dos mesmos parâmetros. Os animais de maior porte (26-37kg) apresentaram maior variância nas medidas de duração e amplitude de onda P e duração do complexo QRS em comparação aos cães de médio (14-25kg) e pequeno (1-13kg) porte. O achado de diferenças entre os sistemas testados deve ser levado em consideração ao se empregar os diversos equipamentos para diagnóstico por meio de ECG-C na rotina clínica, de modo a evitarem-se divergências na interpretação dos exames entre diferentes prestadores de serviços veterinários

Pesquisa Veterinária Brasileira

p. 949-953O método de eletrocardiografia computadorizada (ECG-C) vem sendo crescentemente difundido na medicina veterinária, havendo atualmente diversas marcas e modelos de eletrocardiógrafos disponíveis no mercado. Diante da possibilidade de diferenças na sensibilidade e na reprodutibilidade das medidas obtidas nos traçados, o presente estudo teve como objetivo comparar os parâmetros eletrocardiográficos de cães, obtidos por dois sistemas. Foram avaliados dois diferentes softwares computadorizados, o Wincardio Micromed® (WIN) e o modelo TEB ECGPC® (TEB). Quarenta e dois cães hígidos, de diferentes raças (Cocker Spaniel, Daschund, Labrador, Pinscher, Pit Bull Terrier Poodle, Schnauzer, Shit Tzu, Yorkshire e sem raça definida), machos e fêmeas e com idade entre 4 meses e 16 anos foram agrupados segundo o peso e examinados pelos dois sistemas. As medidas eletrocardiográficas dos diferentes traçados foram analisadas na derivação DII. Os resultados indicaram que o sistema TEB apresentou maior sensibilidade na obtenção das medidas de duração da onda P e do complexo QRS, enquanto o sistema WIN foi mais sensível para determinar as medidas de amplitude dos mesmos parâmetros. Os animais de maior porte (26-37kg) apresentaram maior variância nas medidas de duração e amplitude de onda P e duração do complexo QRS em comparação aos cães de médio (14-25kg) e pequeno (1-13kg) porte. O achado de diferenças entre os sistemas testados deve ser levado em consideração ao se empregar os diversos equipamentos para diagnóstico por meio de ECG-C na rotina clínica, de modo a evitarem-se divergências na interpretação dos exames entre diferentes prestadores de serviços veterinários

Research in Veterinary Science

Texto completo: acesso restrito. p. 340-346An indirect ELISA test was developed for the diagnosis of Brucella canis infection in dogs. A bacterial whole cell extract was used as a solid phase antigen, using B. canis isolated from an infected animal. Sera from culture-positive and healthy negative animals were used as internal reference controls. The cut-off point was determined by a mathematical formula for a statistically valid value, which defined the upper prediction limit, based on the upper tail of the t-distribution of 21 negative control sera readings, for the confidence level of 99.5%. The sensitivity and specificity of the ELISA test were 95% and 91%, respectively. The ELISA test showed a significant concordance index (K = 0.84) with the agar gel immunodiffusion test. The reliability of the ELISA for the detection of infected animals was established by a double blind study testing 280 sera provided by serum banks from different diagnostic and research institutions and analyzed by ROC Curve

Leishmania infantum-derived lipophosphoglycan as an antigen in the accurate serodiagnosis of canine leishmaniasis.

Lipophosphoglycan (LPG) is the major surface glycoconjugate of Leishmania protozoan and has an important biological role in host-parasite interactions both in the midgut epithelium of the sand fly vector and in the vertebrate macrophages. Canine leishmaniasis (CanL) is a chronic infectious disease predominantly caused by Leishmania infantum. An early and accurate immunodiagnosis of the disease is crucial for veterinary clinical practice and for disease control. In this work, we evaluated L. infantum LPG as an antigen in an indirect enzyme-linked immunosorbent assay (ELISA) for CanL immunodiagnosis (LPG-ELISA) by testing serum samples from 97 naturally infected dogs with diverse clinical presentations ranging from subclinical infection to severe disease, as evaluated by veterinarian infectologists. Serum samples from healthy dogs from non-endemic areas (n = 68) and from dogs with other infectious diseases (n = 64) were used as controls for assay validation. The performance of the LPG-ELISA was compared with that of an ELISA using the soluble fraction of L. infantum total lysate antigen (TLA). LPG-ELISA presented a superior performance in comparison to TLA-ELISA, with 91.5% sensitivity, 98.5% specificity and 99.7% accuracy. A distinguishing feature of the LPG-ELISA compared to the TLA-ELISA was its higher ability to identify subclinical infection in clinically healthy dogs, in addition to the absence of cross-reactivity with other canine infectious diseases. Finally, LPG-ELISA was compared to TR DPP visceral canine leishmaniasis test, the immunochromatographic test recommended by the Brazilian Ministry of Agriculture. LPG-ELISA exhibited higher values of specificity (98.5% versus 93.1%) and sensitivity (91.5% versus 90.6%) compared to TR DPP. In conclusion, L. infantum-derived LPG was recognized by antibodies elicited during CanL in different infection stages and was shown to be a suitable antigen for specific clinical settings of veterinary diagnosis and for public health usage