Lactic acid bacterial colonization and human rotavirus infection influence distribution and frequencies of monocytes/macrophages and dendritic cells in neonatal gnotobiotic pigs

Despite accumulating knowledge of porcine macrophages and dendritic cells (DCs) from in vitro studies, information regarding monocytes/macrophages and DCs in lymphoid tissues of enteric pathogen-infected neonatal animals in vivo is limited. In this study we evaluated the influence of commensal bacterial [two strains of lactic acid bacteria (LAB), Lactobacillus acidophilus and L. reuteri] colonization and rotavirus infection on distribution and frequencies of monocytes/macrophages and conventional DCs (cDCs) in ileum, spleen and blood. Gnotobiotic pigs were inoculated with LAB and virulent Wa strain human rotavirus (HRV) (LAB+HRV+), HRV only (LAB-HRV+), LAB only (LAB+HRV-) or mock (LAB-HRV-). The cDCs were characterized as SWC3(+)CD11R1(+), whereas monocytes/macrophages were identified as SWC3(+)CD11R1(-) by flow cytometry in the gnotobiotic pigs at 10 days of age. Infection with HRV alone activated/recruited significantly more monocytes/macrophages to the intestine than LAB colonization and 56% versus 28% of these cells expressed CD14. Colonization with LAB alone also significantly increased the frequencies of monocytes/macrophages and cDCs and the CD14 expression on monocytes/macrophages in ileum and spleen compared to the controls. LAB colonization plus HRV infection significantly reduced macrophage and cDC frequencies in spleen compared to LAB colonization or HRV infection alone, suggesting that LAB colonization down-regulated HRV- infection-induced monocyte/macrophage activation/recruitment at the systemic lymphoid tissue. These results illustrated the distribution of porcine monocytes/macrophages and cDCs and the frequencies of CD14 expression on these cells in intestinal and systemic lymphoid tissues in the early stage of immune responses to intestinal colonization by LAB versus infection by an enteric pathogen HRV and will facilitate further in vivo studies on functional characterization of these immune cells in neonates.     

Strategies of the control of an outbreak of leptospiral infection in dairy cattle in Northeastern Brazil

The aim of the present study was to describe the strategies of the control of an outbreak of leptospiral infection in dairy cattle in Maranhão State, Northeastern Brazil. In the period from January to July 2015, 18 (17%) out of 106 cows presented abortion, six (5.7%) stillbirth, and 12 (11.3%) repeated estrus, totaling 24 animals with reproductive problems. The diagnosis of leptospirosis was based on serology (microscopic agglutination test—MAT), bacteriological culture, and polymerase chain reaction (PCR). Antibiotic therapy, vaccination protocols, and changes in management practices were suggested as control measures. Of all animals on the farm (n?=?280), 136 (48.6%) were seropositive for at least one serovar of Leptospira sp. No pure leptospiral culture was obtained. Eight of the animals with reproductive problems yielded positive PCR results (vaginal fluid of seven animals and urine and vaginal fluid of one animal). Genetic sequencing of a vaginal fluid/urine PCR-positive sample revealed Leptospira borgpetersenii. One year after the adoption of control measures, no reproductive problems were observed. Thus, leptospirosis probably caused the reproductive failures in the herd, and the control and prevention measures implemented were efficient in controlling the disease.

     

Haematological and histopathological findings after ovariectomy in Trypanosoma cruzi infected mice

The aim of this study was to investigate the influences of ovariectomy on histopathological and hematological parameters during the course of Trypanosoma cruzi infection. Hematological and immunological homeostasis is influenced by gonadal steroid hormones. Ovariectomy exerts profound influences on parasitic diseases including T. cruzi infection through modulation of the host's immune response. Three groups of female Mus musculus were infected with 4000 blood trypomastigotes of the Y strain of T. cruzi. One group was subjected to ovariectomy, another to simulated surgery before the infection, and a third group of unoperated animals were used as controls. Marked differences were detected in the responses of blood and tissue parasites. On day 9, post-infection parasitism was significantly higher in ovariectomized animals (P<0.05). These results were confirmed by histopathological studies, in which ovariectomized animals displayed hearts with higher number of amastigote burdens, increased inflammatory infiltrate, enhanced tissue fibers disorganization and decreased lytic antibody percentage, when compared to their counterparts. On day 9 the hematological changes were more apparent, with a decrease in erythrocytes, platelets and leucocytes for ovariectomized infected animals. Simulated surgery, as a stressful agent, did not cause any imbalance in parasitism or in the hemogram profile. The results confirm the importance of the female steroids in resistance against T. cruzi infection.     

Effects of Marine Toxins on the Reproduction and Early Stages Development of Aquatic Organisms

Marine organisms, and specially phytoplankton species, are able to produce a diverse array of toxic compounds that are not yet fully understood in terms of their main targets and biological function. Toxins such as saxitoxins, tetrodotoxin, palytoxin, nodularin, okadaic acid, domoic acid, may be produced in large amounts by dinoflagellates, cyanobacteria, bacteria and diatoms and accumulate in vectors that transfer the toxin along food chains. These may affect top predator organisms, including human populations, leading in some cases to death. Nevertheless, these toxins may also affect the reproduction of aquatic organisms that may be in contact with the toxins, either by decreasing the amount or quality of gametes or by affecting embryonic development. Adults of some species may be insensitive to toxins but early stages are more prone to intoxication because they lack effective enzymatic systems to detoxify the toxins and are more exposed to the toxins due to a higher metabolic growth rate. In this paper we review the current knowledge on the effects of some of the most common marine toxins on the reproduction and development of early stages of some organisms.     

Life cycle inference and phylogeny of Ortholinea labracis n. sp. (Myxosporea: Ortholineidae), a parasite of the European seabass Dicentrarchus labrax (Teleostei: Moronidae), in a Portuguese fish farm

Ortholinea labracis n. sp. is described and its life cycle is inferred from a Southern Portuguese fish farm, with basis on microscopic and molecular procedures. This myxosporean parasite infects the urinary bladder of the European seabass Dicentrarchus labrax and the intestinal epithelium of a marine oligochaete of the genus Tectidrilus. Myxospores subspherical in valvular view and ellipsoidal in sutural view measuring 7.6 ± 0.3 (6.8–8.7) μm in length, 7.2 ± 0.2 (6.7–7.7) μm in width and 6.5 ± 0.4 (5.8–7.7) μm in thickness. Two polar capsules, 3.0 ± 0.2 (2.6–3.4) μm long and 2.4 ± 0.1 (2.0–2.9) μm wide, located at the same level, but with divergent orientation and opening to opposite sides of the suture line. Sequencing of the SSU rRNA gene revealed a similarity of 100% between the analysed myxospores and triactinomyxon actinospores. The phylogenetic setting of O. labracis n. sp. shows subgrouping in correlation with tissue tropism, but identifies this parasite as another exception to the main division of Myxosporea into the main freshwater and marine lineages.     

Cylindrospermopsin effects on cell viability and redox milieu of Neotropical fish <Emphasis Type="Italic">Hoplias malabaricus</Emphasis> hepatocytes

Cylindrospermopsin (CYN) is a cyanotoxin that is cytotoxic to a wide variety of cells, particularly to the hepatocytes. In this study, the toxic effects of purified CYN were investigated in primary cultured hepatocytes of Neotropical fish Hoplias malabaricus. After isolation, attachment, and recovery for 72 h, the cells were exposed for 72 h to 0, 0.1, 1.0, 10, and 100 μg l^?1 of CYN. Then, cell viability and a set of oxidative stress biomarker responses were determined. Catalase, superoxide dismutase, glucose-6-phosphate dehydrogenase, and glutathione S-transferase activities were not affected by exposure to CYN. Concentration-dependent decrease of glutathione reductase activity occurred for most CYN-exposed groups, whereas non-protein thiol content increased only for the highest CYN concentration. Lipid peroxidation, protein carbonylation, and DNA damage levels were not altered, but reactive oxygen species levels increased in the cells exposed to the highest concentration of CYN. Cell viability decreased in all the groups exposed to CYN. Thus, CYN may cause a slight change in redox balance, but it is not the main cause of cell death in H. malabaricus hepatocytes.     

First description of a natural infection with spleen and kidney necrosis virus in zebrafish

Zebrafish has become a popular research model in the last years, and several diseases affecting zebrafish research facilities have been reported. However, only one case of naturally occurring viral infections was described for this species. In 2015, infectious spleen and kidney necrosis virus (ISKNV) was detected in zebrafish from a research facility in Spain. Affected fish showed lethargy, loss of appetite, abnormal swimming, distention of the coelomic cavity and, in the most severe cases, respiratory distress, pale gills and petechial haemorrhages at the base of fins. Cytomegaly was the most relevant histopathological finding in organs and tissues, sometimes associated to degenerative and necrotic changes. ISKNV belongs to the relatively newly defined genus Megalocytivirus, family Iridoviridae, comprising large, icosahedral cytoplasmic DNA viruses. This is the first case of naturally occurring Megalocytivirus infection in zebrafish research facilities, associated with morbidity. The virus has been identified based on both pathologic and genetic evidence, to better understand the pathogenesis of the infection in zebrafish and the phylogenetic relationship with other iridoviruses. Given the ability of megalocytiviruses to cross‐species boundaries, it seems necessary to implement stringent biosecurity practices as these infections may invalidate experimental data and have major impact on laboratory and cultured fish.     

Blueberry Residue Encapsulation by Ionotropic Gelation

In the processing of fruits such as blueberry (Vaccinium sp), that has high levels of phenolic acid, the food industry produces tons of organic waste that causes harm to the environment. Encapsulation is a technique used to take advantage of these wastes. Several methods are used to encapsulate substances, among them ionotropic gelation proves to be a simple, precise, efficient and economical method for obtaining particles with encapsulated bioactives. In this manner, the aim of this study was to test sodium alginate as wall material to encapsulate blueberry residue by ionotropic gelation. The microbeads were characterized by scanning electron microscopy (SEM), x-ray diffraction (XRD), total phenolic compounds, antioxidant capacity and in vitro dissolution. The results showed that the microbeads had surface invagination; retention of 67.01% of the phenolic compounds after encapsulation and 68.2%, phenolic release 120 min after in vitro dissolution. The results suggest that the tested matrix was suitable for encapsulation. The produced microbeads are promising for applications in food products, once the phenolic compounds present in the blueberry residues were maintained after encapsulation.     

Ecophysiology of iron homeostasis in plants

In nature, iron (Fe) occurs in abundance and ranks fourth among all elements on Earth’s surface. Still, its availability to plants is reduced, once this element is in the form of hydrated oxides, which can limit plant productivity and biomass production. On the other hand, in high concentrations, this essential micronutrient for the plants can become a toxic agent, increasing the environmental contamination. Fe is necessary for the maintenance of essential processes like respiration and photosynthesis, participating in the electron transport chain and in the conversion between Fe²⁺ and Fe³⁺, being a key element for carbon dioxide (CO₂) fixation and, therefore, important for crop production of cultivated or natural species. The balance of Fe should be strictly controlled, because both its deficiency and its toxicity affect the physiological process of plants. In aerated soils Fe is present in the form of Fe³⁺, which is the oxidized form and is less available to plants, so these organisms have developed different strategies for absorption, transport and storage of Fe. Deficiency and excess of Fe correlate with local soil conditions and with the care adopted in plant nutrition during the phenological phases and/or in the course of its cultivation. In situations of excessive accumulation of Fe in tissues, an enhancement of hydroxyl radical generation (OH^?) occurs by Fenton reaction. Here, we review the nutritional, genetic and ecophysiological aspects of uptake, translocation and accumulation of Fe ions in plants growing under conditions of deficiency or toxicity of this metal.     

Epitypification,characterization and phylogenetic positioning of Pseudobeltrania cedrelae,the causal agent of pseudobeltrania spot on Cedrela fissilis

The fungus Pseudobeltrania cedrelae, the type species of the genus, is the causal agent of an important leaf spot in seedlings and adult plant of cedar (Cedrela fissilis). Due to the contradictory phylogenetic position of the genus Pseudobeltrania, epitypification of P. cedrelae was carried out based on a culture obtained from the same locality and host of the original type. Samples were collected, and 10 isolates of P. cedrelae associated with lesions on cedar leaflets were obtained. For morphological characterization, conidia, conidiophores, conidiogeneous cells, conidiogeneous loci and basal cells were taken both from the fungus obtained from leaf lesions and from that obtained in slide culture. Mycelial growth rates and sporulation were evaluated in six different culture media. For molecular phylogeny, maximum parsimony analyses were performed from the ITS and 28S sequences of the isolates. Both the morphological characteristics of the fungal structures obtained from symptomatic leaves and the slide culture technique presented variations. In foliar lesions, isolates presented the same morphological characteristics as the type material. Mycelial growth rate and sporulation of P. cedrelae were greatest on malt extract agar and V8 juice agar. Pseudobeltrania cedrelae was pathogenic when inoculated into healthy cedar plants. According to the phylogenetic tree, isolates grouped in the same clade, but in a distinct clade of Pseudobeltrania ocoteae. The results suggested that P. ocoteae belongs to the genus Hemibeltrania. This paper presents new information on P. cedrelae that contributes to clarifying the phylogeny of the Beltraniaceae.