RESUMO
Acute phase proteins have been used as tools for the diagnosis, monitoring, and prognosis of several diseases in domestic animals. However, the dynamics of these proteins in infection by Trypanosoma cruzi, the causative agent of Chagas disease in dogs, is still unknown. The aim of this study was to determine concentrations of acute phase proteins (C-reactive protein, haptoglobin, ferritin and paraoxonase-1) in dogs in a coastal town of Ecuador, with natural Trypanosoma cruzi infection with or without seroreactivity of Ehrlichia canis, Ehrlichia ewingii, Anaplasma phagocytophilum, Anaplasma platys, Borrelia burgdorferi and Dirofilaria immitis. For the detection of Trypanosoma cruzi serum antibodies, two different antigen-based enzyme-linked immunosorbent assay tests were implemented. For the detection of seroreactivity of Ehrlichia canis, Ehrlichia ewingii, Anaplasma phagocytophilum, Anaplasma platys, Borrelia burgdorferi and Dirofilaria immitis, an IDEXX SNAP® 4Dx® test was used. To determine the concentration of C-reactive protein and ferritin, an immunoturbidimetric assay was used; haptoglobin concentration was measured using a commercial colorimetric method validated in dogs; a spectrophotometric method was used to determine the serum concentration of paraoxonase-1. Results showed a reduction in the serum levels of paraoxonase-1 in Trypanosoma cruzi-seroreactive dogs, either with or without seroreactivity to other vector-borne diseases. A serum ferritin increment was observed in Trypanosoma cruzi-seroreactive dogs with seroreactivity to any other vector-borne diseases. Our findings suggest that paraoxonase-1 levels are reduced in Trypanosoma cruzi-seroreactive dogs without evident clinical signs of Chagas disease, despite their seroreactivity to the other vector-borne diseases studied. These findings could indicate an oxidative stress response in Trypanosoma cruzi-seroreactive dogs with no evident signs of inflammation.
RESUMO
Dogs are a reservoir for Chagas disease, caused by Trypanosoma cruzi (T. cruzi), and other companion vector-borne diseases, including ehrlichiosis (Ehrlichia canis and Ehrlichia ewingii), anaplasmosis (Anaplasma phagocytophilum and Anaplasma platys), dirofilariasis (Dirofilaria immitis) and Lyme disease (Borrelia burgdorferi). This study has two key objectives: 1) to determine seroreactivity against T. cruzi in dogs from the town of Colón, in Portoviejo city, in the central coast of Ecuador; and 2) to establish the coinfection frequency of other companion vector-borne diseases in dogs positive for T. cruzi. Antibodies against T. cruzi were detected using two enzyme-linked immunosorbent assays. Diagnostic consensus between ELISA tests was established using the Cohen's Kappa coefficient. Other haemoparasitic diseases were detected using the IDEXX SNAP® 4Dx® kit in dogs previously diagnosed as T. cruzi-seropositive. From 84 dogs sampled, 57.14% (48/84) tested positive for T. cruzi. Co-infection analysis of 25 dogs positive for T. cruzi revealed antibodies also against Ehrlichia spp. (48%), Anaplasma spp. (28%), and Dirofilaria immitis (12%). These results provide a novel perspective regarding the status of these pathogens which co-infect dogs in Colón. Since all these pathogens are zoonotic, our findings should warn regional health authorities to implement sanitary programs, to better prevent and control vectors associated to these pathogens. On the other hand, human and veterinarian doctors, should consider that patients with a cardiac infection condition could be suffering co-infections with two or more vector transmitted pathogens.
Assuntos
Anaplasmose , Borrelia burgdorferi , Doença de Chagas , Coinfecção , Doenças do Cão , Ehrlichiose , Doença de Lyme , Trypanosoma cruzi , Doenças Transmitidas por Vetores , Anaplasma , Anaplasmose/epidemiologia , Animais , Anticorpos Antibacterianos , Doença de Chagas/epidemiologia , Doença de Chagas/veterinária , Coinfecção/epidemiologia , Coinfecção/veterinária , Doenças do Cão/epidemiologia , Cães , Equador/epidemiologia , Ehrlichiose/epidemiologia , Ehrlichiose/veterinária , Humanos , Estudos SoroepidemiológicosRESUMO
BACKGROUND: Leptin has been measured in human in saliva samples. However, the low leptin concentration found in this biological fluid makes necessary the use of high sensitive methods. To the authors' knowledge, leptin has not been measured in porcine saliva. This study aimed to develop and validate a time-resolved immunofluorometric assay (TR-IFMA) for salivary leptin measurements in pigs, using a species-specific antibody, and to evaluate how salivary leptin changes with body weight, food ingestion, and in experimental models of stress and inflammation. Polyclonal antibodies were produced in rabbits immunized with recombinant porcine leptin and used to develop a sandwich TR-IFMA. RESULTS: The method had intra-assay and inter-assay coefficients of variation lower than 10 and 16 %, respectively. The assay was accurate and the low limit of detection allowed detection of leptin in all analyzed samples. Salivary leptin concentration was positively correlated to body weight (r = 0.58, P = 0.01) and increased after food ingestion (P < 0.001) and after 24 h of applying a model of experimental inflammation by turpentine injection (P < 0.05). However, it did not significantly change after a model of acute stress consisting of a nose snare restraining. CONCLUSION: These results indicate that the developed assay can measure leptin in porcine saliva in a reliable way and that leptin in saliva is influenced by body weight, food ingestion and inflammation.
Assuntos
Fluorimunoensaio/veterinária , Leptina/análise , Saliva/química , Suínos , Animais , Western Blotting/veterinária , Fluorimunoensaio/métodos , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Ancylostoma spp. is one of the most prevalent canine intestinal nematode infections which usually causes subclinical disease in adult dogs and has zoonotic implications. Therefore, the aim of this study was to explore and evaluate the possible pathophysiological changes that Ancylostoma spp. could produce in female dogs naturally infected but without clinical signs of disease, by screening a wide variety of biochemical markers for potential changes. Samples of feces and blood of 45 dogs were collected and fecal flotation and zinc sulphate centrifugal flotation were performed. The biochemical analytes determined were: the acute-phase proteins C-reactive protein (CRP) and haptoglobin (Hp); the lipid profile (cholesterol, triglycerides, HDL, LDL); the serum iron profile: iron, unsaturated iron binding-capacity (UIBC), and ferritin; the enzyme butyrylcholinesterase (BChe); the pancreatic profile: amylase, lipase, and trypsin-like immunoreactivity (TLI); the oxidative stress markers: total antioxidant capacity (TAC) and paraoxonase -1 (PON-1), along with total protein, albumin, and insulin-like growth factor - 1 (IGF - 1). Ancylostoma spp. eggs were detected in 29/45 dogs (64.4 %). Dogs were divided into two groups according to the results of fecal flotation methods. Group 1: negative fecal floatation (n = 16), and Group 2: subclinical infection with the observation of Ancylostoma spp. type eggs/x 40 objective fields (n = 29). RESULTS: Mann-Whitney U test was used to compare the biochemical analyte results between the two groups (P < 0.05). Significant increases in CRP (µg/mL) (median): non-infected dogs: 5.5; subclinically infected dogs 18.7; P = 0.03, Hp (g/L) (median): G1: 2.4; G2: 3.3; P = 0.03, and UIBC (µg/dL) (median): non-infected dogs: 139.4; subclinically infected dogs: 216; P = 0.0015, and significantly decreased iron (µg/dL) (median): non-infected dogs: 202.5; subclinically infected dogs: 125.7; P = 0.0041, IGF-1 (ng/mL) (median): non-infected dogs: 224; subclinically infected dogs: 123; P = 0.02, and albumin (g/dL) (median): non-infected dogs: 2.8; subclinically infected dogs: 2.5; P = 0.04 concentrations were observed in dogs with subclinical Ancylostoma spp. infection when compared to non-infected dogs. CONCLUSION: These findings provide an overview of the biochemical effects produced by patent Ancylostoma spp. in naturally infected dogs without any evident clinical signs of disease, which could be considered in differential diagnosis, especially in an endemic area for this parasite.
Assuntos
Ancylostoma , Ancilostomíase/veterinária , Doenças do Cão/parasitologia , Ancilostomíase/sangue , Animais , Doenças do Cão/sangue , Cães , Fezes/parasitologia , Feminino , ÓvuloRESUMO
Eleven Murciano-Granadina goats in late pregnancy were separated into two groups (1) control (n=6) and (2) fasting for 72 h to induce pregnancy toxaemia (n=5). Venous blood was taken daily to determine acid-base and electrolyte parameters. Significant decreases in blood pH, bicarbonate concentration and base excess, and a significant increase in anion gap were observed after 24h of fasting. These changes were significantly correlated with non-esterified fatty acid concentration. No significant changes were observed in pCO(2), and electrolyte or lactate concentrations. Clinical signs of pregnancy toxaemia in fasted goats appeared by 72h post-fasting. These signs and the changes in acid-base balance disappeared once feed was reintroduced. Blood pH, bicarbonate concentration, base excess and anion gap could be indicators of early pregnancy toxaemia in goats.
Assuntos
Desequilíbrio Ácido-Base/veterinária , Doenças das Cabras/fisiopatologia , Toxemia/veterinária , Desequilíbrio Hidroeletrolítico/veterinária , Equilíbrio Ácido-Base , Desequilíbrio Ácido-Base/sangue , Desequilíbrio Ácido-Base/etiologia , Desequilíbrio Ácido-Base/fisiopatologia , Animais , Bicarbonatos/sangue , Análise Química do Sangue/veterinária , Feminino , Doenças das Cabras/sangue , Doenças das Cabras/etiologia , Cabras , Concentração de Íons de Hidrogênio , Gravidez , Distribuição Aleatória , Toxemia/sangue , Toxemia/complicações , Toxemia/fisiopatologia , Desequilíbrio Hidroeletrolítico/sangue , Desequilíbrio Hidroeletrolítico/etiologia , Desequilíbrio Hidroeletrolítico/fisiopatologiaRESUMO
The present work aimed to study the behavior of acute phase proteins (haptoglobin, serum amyloid A, acid soluble glycoprotein, fibrinogen, and albumin) in fasting-induced pregnancy toxemia in goats and their relationship with classical indicators of this disorder such as beta-hydroxybutyrate and nonesterified fatty acids in the blood and decreased urine pH and ketonuria. Twelve adult Murciano-Granadina goats at the final stage of gestation were used in this experiment. Pregnancy toxemia was induced in 6 goats by fasting for 72 hr. The other 6 animals were used as control group. Ketonuria was present in 4 out of 5 fasting animals at 24 hr and in all fasting animals at 48 hr of fasting. Serum nonesterified fatty acids were significantly increased at 24, 48, and 72 hr of fasting. Beta-hydroxybutyrate and haptoglobin achieved significantly increased concentrations at 48 hr and 72 hr, respectively, remaining increased during the entire study. Serum amyloid A, acid soluble glycoprotein, fibrinogen, and albumin were not affected by fasting. In conclusion, acute phase proteins (including haptoglobin) seemed not to have an advantage over traditional markers in diagnosis of fasting-induced pregnancy toxemia in goats.
Assuntos
Doenças das Cabras/metabolismo , Pré-Eclâmpsia/veterinária , Animais , Feminino , Fibrinogênio/análise , Fibrinogênio/metabolismo , Cabras , Haptoglobinas/análise , Haptoglobinas/metabolismo , Pré-Eclâmpsia/metabolismo , Gravidez , Distribuição Aleatória , Albumina Sérica/análise , Albumina Sérica/metabolismo , Proteína Amiloide A Sérica/análise , Proteína Amiloide A Sérica/metabolismo , Estatísticas não ParamétricasRESUMO
Acute phase proteins (APPs) are important diagnostic indicators of inflammatory disturbances in animals. The objectives of the current study were to validate analytical methods for measuring haptoglobin (Hp), serum amyloid A (SAA), acid soluble glycoprotein (ASG), fibrinogen, and albumin concentrations in goats and to evaluate their response to an inflammatory stimulus in this species. Intra- and interassay coefficients of variation (CVs) were in the range 0.07-9.31% and 1.83-12.68%, respectively, for all APPs and showed good precision. All assays determined APPs in a linear manner at different sample dilutions with high correlation coefficients with the exception of fibrinogen, which was measured by the heat precipitation method. Subcutaneous injection of turpentine oil induced an increase in Hp, SAA, ASG, and fibrinogen serum concentrations and a decrease in albumin concentration.