A new cyclodipeptide from Tetragonisca angustula honey active against Neospora caninum and in silico study.

A new cyclic natural compound formed by succinic acid and two alanine amino acid units was isolated from the Tetragonisca angustula honey extract. The chemical structure of 1 was established based on spectroscopic data analysis, including one- (1H and 13C NMR) and two-dimensional NMR techniques (1H-1H-COSY, HSQC and HMBC). A primary culture model previously infected with Neospora caninum was used to evaluate 1 for two time intervals (24 and 72 h), showing a reduction (40-56%) of the number of tachyzoites in the first 24 h and until 72 h, a dose-dependent reduction in parasite proliferation (25-50%). Glial cells treated with 1 did not demonstrate toxicity at concentrations up to 25 ug/mL. Treated and infected cultures showed an increase in NO when compared to control cells in 24 h and 72 h. In silico studies suggest that the new compound may affect DNA synthesis and impair -protein production.

Activation of the Kynurenine Pathway and Production of Inflammatory Cytokines by Astrocytes and Microglia Infected With Neospora caninum.

In the central nervous system, astrocytes and microglia contribute to homeostasis, regulating the immune response to infectious agents. Neospora caninum is an obligate intracellular protozoan that infects different animal species and it is encysted in their nervous tissue while triggering an immune response modulated by glia. This study aimed to evaluate the infection of primary cultures of rat glial cells by N. caninum through the catabolites of tryptophan, the expression of inflammatory mediators and the integrity of neural tissue. Infection with this coccidium resulted in morphological and functional changes, particularly astrogliosis and microgliosis, and increased the expression of the inflammatory mediators TNF, IL1ß, IL-10, and arginase, as well as mRNA for CCL5 and CCL2, molecules involved in the CNS chemotaxis. The infection with N. caninum in glial cells also triggered the activation of the tryptophan pathway, characterized by increased kynurenine 2,3 monooxygenase (KMO) mRNA expression, and by the production of the excitotoxin quinolinic acid (QUIN). Moreover, glia-neuron co-cultures, when exposed to the secretome derived from N. caninum infected glial cells, presented greater neurons distribution and formation of neurite extensions, associated to morphological changes in astrocytes compatible with neuro-preservation. Considering that the tryptophan catabolism is associated to immune response, these findings suggest that glial activation in N. caninum infection should be responsible for modulating the inflammatory status in an attempt to restore the nervous system homeostasis, since excessive inflammatory response can cause irreversible damage to tissue preservation.

Flaxseed added to the diet of Alpine goats affects the nutrients intake and blood parameters.

The flaxseed is a nutraceutical food used as a source of α-linolenic acid, which can bring benefits to the health of mammals. This study was carried out to examine the effect of flaxseed inclusion in the diets on the intake of nutrients, body weight, and blood parameters of Alpine goats. Twenty-one adult females with an initial average weight of 41.06 ± 1.84 kg were used in a completely randomized design, with four experimental treatments (0, 5, 10, and 15% of flaxseed in the total diet) and five replications per treatment. The intake of the dry matter presented a decreasing linear effect (P < 0.001), with a reduction of 53.5% between the control diet (0% of flaxseed) and 15% of flaxseed, but no effect was observed on weight gain (P > 0.05). Inclusion of flaxseed from 0 to 15% linearly decreased the intakes of organic matter, crude protein, non-fibrous carbohydrates, and neutral detergent fiber, but increased the ether extract intake (P < 0.001). Regarding plasma concentration traits, increasing the flaxseed levels from 0 to 15% had linear positive effect on LDL, VLDL, and triglycerides (P < 0.05), but no effect on cholesterol, HDL, glucose, creatinine, and urea (P > 0.05). The inclusion of flaxseed in the diet for 80 days changes the nutrients intake and blood parameters but shows no impact on body weight. However, further studies are required to determine the impact of flaxseed on the goat's health in the long term since there has been an increase in the concentration of triglycerides, LDL, and VLDL.

Neurological Infection, Kynurenine Pathway, and Parasitic Infection by Neospora caninum.

Neuroinflammation is one of the most frequently studied topics of neurosciences as it is a common feature in almost all neurological disorders. Although the primary function of neuroinflammation is to protect the nervous system from an insult, the complex and sequential response of activated glial cells can lead to neurological damage. Depending on the type of insults and the time post-insult, the inflammatory response can be neuroprotective, neurotoxic, or, depending on the glial cell types, both. There are multiple pathways activated and many bioactive intermediates are released during neuroinflammation. One of the most common one is the kynurenine pathway, catabolizing tryptophan, which is involved in immune regulation, neuroprotection, and neurotoxicity. Different models have been used to study the kynurenine pathway metabolites to understand their involvements in the development and maintenance of the inflammatory processes triggered by infections. Among them, the parasitic infection Neospora caninum could be used as a relevant model to study the role of the kynurenine pathway in the neuroinflammatory response and the subset of cells involved.

Role of IFN-γ and LPS on neuron/glial co-cultures infected by Neospora caninum.

Neospora caninum causes cattle abortion and neurological symptoms in dogs. Although infection is usually asymptomatic, classical neurological symptoms of neosporosis may be associated with encephalitis. This parasite can grow in brain endothelial cells without markedly damages, but it can modulate the cellular environment to promote its survival in the brain. In previous studies, we described that IFN-γ decreased the parasite proliferation and down regulated nitric oxide (NO) production in astrocyte/microglia cultures. However, it remains unclear how glial cells respond to N. caninum in the presence of neurons. Therefore, we evaluated the effect of 300 IU/mL IFN-γ or 1.0 mg/mL of LPS on infected rat neuron/glial co-cultures. After 72 h of infection, LPS did not affect the mitochondrial dehydrogenase activity. However, IFN-γ decreased this parameter by 15.5 and 12.0% in uninfected and infected cells, respectively. The number of tachyzoites decreased 54.1 and 44.3% in cells stimulated with IFN-γ and LPS, respectively. Infection or LPS treatment did not change NO production. On the other hand, IFN-γ induced increased nitrite release in 55.7%, but the infection reverted this induction. IL-10 levels increased only in infected cultures (treated or not), meanwhile PGE2 release was improved in IFN-γ/infected or LPS/infected cells. Although IFN-γ significantly reduced the neurite length in uninfected cultures (42.64%; p < 0.001), this inflammatory cytokine reverted the impairment of neurite outgrowth induced by the infection (81.39%). The results suggest a neuroprotective potential response of glia to N. caninum infection under IFN-γ stimulus. This observation contributes to understand the immune mediated mechanisms of neosporosis in central nervous system (CNS).

Loss of infectivity of Neospora caninum oocysts maintained for a prolonged time.

The purpose of this study was to investigate whether sporulated Neospora caninum oocysts, which had been stored for 46 mo in a 2% sulfuric acid solution at 4 degrees C, remain morphologically viable and infective to gerbils (Meriones unguiculatus). Six gerbils were orally inoculated with doses of 400 or 1,200 oocysts. Two mo after inoculation, the animals did not show any clinical signs, had no histological lesions, and were seronegative for N. caninum at 1: 50 in an immunofluorescent antibody test. PCR using the brain from each gerbil did not reveal N. caninum specific DNA. We conclude that oocysts preserved for 46 mo are not infective, despite being morphologically intact.

Frequência de anticorpos anti-Neospora caninum em cães nos municípios de Salvador e Lauro de Freitas, Estado da Bahia - Brasil / Frequency of antibodies anti-Neospora caninum on dogs from Salvador and Lauro de Freitas, Bahia state - Brasil

A freqüência de anticorpos IgG anti-Neospora caninum foi estudada em 415 amostras séricas de cães domiciliados e errantes, procedentes dos municípios baianos de Salvador e Lauro de Freitas, utilizando-se a técnica de imunofluorescência indireta, com ponto de corte igual a 1:50. Anticorpos da classe IgG anti-N. caninum foram detectados em 13,3% (22/165) dos cães domiciliados e em 11,2% (28/250) dos errantes. As freqüências de soropositivos machos e fêmeas foram 8,0% (6/75) e 18,4% (14/76) nos cães domiciliados e 12,6% (17/135) e 9,6% (11/115) nos errantes, respectivamente. Não houve diferenças estatisticamente significativas entre sexo, idade, raça e a freqüência de soropositividade ao N. caninum dos cães domiciliados e errantes.

The frequency of anti-Neospora caninum IgG antibodies was studied in 415 serum samples from owned and stray dogs from Salvador and Lauro de Freitas counties. The Indirect immunofluorescence was performed using a cut-off of 1:50. Anti-N. caninum IgG antibodies were detected in 13.3% (22/165) owned dogs and in 11.2% (28/250) stray dogs. The frequencies of males and females seropositivity were 8.0% (6/75) and 18.4% (14/76) for owned dogs and 12.6 % (17/ 135) and 9.6% (11/115) in the stray dogs population, respectively. There were not a statistical significant difference between sex, age, breed and the dogs anti-N. caninum IgG antibodies incidence.