Sheep naturally infected with Anaplasma ovis - evaluation of cardiac and inflammatory biomarkers

Background: Anaplasmosis, also called gall sickness or tropical bovine ehrlichiosis, is an infectious disease caused by species belonging to the genus Anaplasma in domestic and wild animals in tropical and subtropical regions. Anaplasma ovis and A. phagocytophilum are important pathogens of sheep. A. ovis is considered the most common species affecting sheep. The infection is usually subclinical and progresses with high fever, anaemia, icterus, weight loss and abortions. This study aimed to investigate changes in cardiac damage markers, oxidative stress and antioxidant status, cytokines, and acute phase proteins in sheep naturally infected with A. ovis. Materials, Methods & Results: For this purpose, a total of 40 animals, including 20 healthy sheep and 20 sheep infected with anaplasmosis, were used. A. ovis was diagnosed based on clinical findings and peripheral blood smear. Blood smears were prepared from the ear vein. The smears were stained with Giemsa and examined for the presence of Anaplasma spp. Infection was also confirmed by polymerase chain reaction (PCR) analysis. The genomic DNA was isolated from blood, and the MSP-4 gene region was amplified as A. ovis specific target gene. Twenty clinically healthy sheep of the same age group, reared under the same conditions and testing negative in the molecular assessment were used as controls. Blood samples were collected from the cephalic vein and and centrifuged to obtain serum. The serum stored at -20°C until the analysis stage. Serum samples were used for the analysis of cardiac damage markers [troponin I (cTnI), creatine kinase MB (CK-MB), lactate dehydrogenase (LDH) and aspartate transaminase (AST)], oxidative stress parameters [malondialdehyde (MDA), total antioxidant status (TAS), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)], cytokines [interleukins IL-6, IL-1ß and IL-10, tumour necrosis factor α (TNF-α), and interferon-γ (IFN-γ)] and acute phase proteins [C-reactive protein (CRP), serum amyloid A (SAA) and haptoglobin (Hp)]. cTnI and CK-MB levels were measured using a chemiluminescent immunoassay. MDA, TAS, SOD, CAT, GPx, TNF-α, IL-1ß, IL-6, IL-10, IFN-γ, SAA and Hp levels were measured by an ELISA reader. LDH, AST and CRP levels were measured in an autoanalyzer. cTnI and LDH levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The concentration of AST was decreased in infected animals. MDA, TAS, SOD, CAT and GPx levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The levels of the inflammatory parameters such as TNF-α, IL-1ß, IL-10 and IFN-γ were significantly increased in the infected animals compared to the healthy ones (P < 0.05). Hp level were significantly increased in the infected group compared with the control group (P < 0.05). However, there was no significant change in CK-MB, SAA and CRP concentrations in the infected animals (P > 0.05). Discussion: Ovine anaplasmosis is an obligate intracellular arthropod disease that causes widespread changes in haematobiochemical, immune response and oxidative stress parameters. Cardiac damage is often overlooked in field conditions due to the lack of adequate knowledge about the pathophysiology of the disease. Our results showed that A. ovis infection leads to significant changes in cardiac biomarkers and that the parasite can cause cardiac dysfunction. This is the first report on cardiac damage markers in Anaplasma-infected sheep. Additionally, the levels of proinflammatory and oxidative stress markers that may cause functional disorders were also found to be increased. Thus, measuring markers of cardiac function, oxidative stress and inflammation can be a useful tool in the early diagnosis of ovine anaplasmosis.

Evaluation of echocardiographic variables of morphometry and function in horses submitted to minimally invasive partial pericardiotomy / [Avaliação de variáveis ecocardiográficas morfométricas e funcionais em cavalos submetidos à pericardiotomia parcial minimamente invasiva]

This study aimed to evaluate the impact of minimally invasive partial pericardiotomy on echocardiographic variables of morphometry and function in healthy horses. Minimally invasive pericardiotomy was performed in six healthy horses. Echocardiographic evaluation was executed in different moments: prior to the surgical procedure (M0); 24 hours post procedure (M1); 72 hours post procedure (M2) and 28 days post procedure (M3). The following variables were measured: Right ventricular internal diameter in diastole and systole (RVd and RVs), interventricular septum thickness in diastole and systole (IVSd and IVSs), left ventricular internal diameter in diastole and systole (LVd and LVs), left ventricular free wall thickness in diastole and systole (LVFWd and LVFWs), aortic root diameter (Ao) and left atrial diameter (LA). From this data, the following variables were calculated: fractional shortening (FS%), fractional thickening of the interventricular septum (IVS%), fractional thickening of the left ventricular free wall (LVFW%) and the relationship between left atrial and aortic diameters (LA/Ao). After 28 days, a new thoracoscopy was performed for inspection of the thoracic cavity. In M1 and M2 ECO evaluations, a statistically significant change in LVFW and a decrease in RVd, LVd, LVFWs, LA, LVs, FS% and IVS was documented. Pericardiotomy is a promising technique in horses, with minor postoperative complication. The variations in the echocardiographic parameters were transient and did not cause hemodynamic damage to the animals.(AU)

O objetivo do presente estudo foi avaliar o impacto da pericardiotomia parcial minimamente invasiva sobre as variáveis ecocardiográficas morfométricas e funcionais em cavalos, visto que não há nenhum trabalho que tenha avaliado o impacto da pericardiotomia na espécie equina. Foram utilizados seis cavalos hígidos, nos quais se realizou pericardiotomia minimamente invasiva. Em todos eles, foi efetuado exame ecocardiográfico em diferentes momentos: previamente ao procedimento cirúrgico (M0); 24 horas após (M1); 72 horas após (M2) e 28 dias após (M3). Foram mensurados: diâmetro interno do ventrículo direito (VDd e VDs), espessura do septo interventricular (SIVd e SIVs), diâmetro interno do ventrículo esquerdo (VEd e VEs), espessura da parede livre do ventrículo esquerdo (PLVEd e PLVEs), diâmetro interno da aorta (Ao) e diâmetro atrial esquerdo (AE). Foram calculadas as variáveis fração de encurtamento (FEC%), espessamento fracional do septo interventricular (SIV%), espessamento fracional da parede livre do ventrículo esquerdo (PLVE%) e relação entre diâmetro do átrio esquerdo e diâmetro aórtico (AE/Ao). Após 28 dias, realizou-se nova toracoscopia para inspeção da cavidade torácica. Nas avaliações do ECO no M1 e no M2, houve alteração estatisticamente significativa no PLVE em diástole (28±5,5 e 31,7±2, respectivamente) e diminuição dos parâmetros VDd,(30,1±11,6 e 31,7± 10,7) VEd (113,3±21 e 121,7±13,7), PLVEs (42±8,2 e 43,9±2,8), AE (78,5±6,1 e 82,7±4,7), VEs (74,1±16 e 71,5±9,3), FEC (34,4±10,2 e 41,2±5,7) e SIV (27,1±8,7 e 42,3±27,9). A técnica de pericardiotomia empregada mostrou-se promissora em equinos. As variações dos parâmetros ecocardiográficos foram transitórias, não causando prejuízos hemodinâmicos aos animais.(AU)