RNAi as an emerging approach to control Fusarium head blight disease and mycotoxin contamination in cereals

Fusarium graminearum is a major fungal pathogen of cereals worldwide, causing seedling, stem base and floral diseases, including Fusarium head blight (FHB). In addition to yield and quality losses, FHB contaminates cereal grain with mycotoxins, including deoxynivalenol, which are harmful to human, animal and ecosystem health. Currently, FHB control is only partially effective due to several intractable problems. RNA interference (RNAi) is a natural mechanism that regulates gene expression. RNAi has been exploited in the development of new genomic tools that allow the targeted silencing of genes of interest in many eukaryotes. Host‐induced gene silencing (HIGS) is a transgenic technology used to silence fungal genes in planta during attempted infection and thereby reduces disease levels. HIGS relies on the host plant's ability to produce mobile small interfering RNA molecules, generated from long double‐stranded RNA, which are complementary to targeted fungal genes. These molecules are transferred from the plant to invading fungi via an uncharacterised mechanism, to cause gene silencing. Here, we describe recent advances in RNAi‐mediated control of plant pathogenic fungi, highlighting the key advantages and disadvantages. We then discuss the developments and implications of combining HIGS with other methods of disease control. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Potato Virus Y NTN : A Coat and P1 Protein Sequences Analysis of a Brazilian Isolate

Potato tubers (Solanum tuberosum L. cv. Monalisa) showing superficial necrotic rings typical of Potato virus Y ^NTN (PVY^NTN) infection were collected from commercial potato fields in the State of São Paulo (Brazil). An isolate named IAC-PVY^NTN was characterized by sequencing 822 and 836 nucleotides from the coat protein and P1 protein coding genes, respectively. The IAC-PVY^NTN was observed as a European/North American recombinant isolate closely related to three Eu-PVY^NTN isolates that share an alternative recombination site in the coat protein cistron near position 144. Therefore, the IAC-PVY^NTN isolate may represent a novel recombinant variant between PVY^N N-605 Swiss (European) and PVY O-139 Canadian (North American). Cluster analysis of the P1 sequences found IAC-PVY^NTN in the sub-cluster of Eu-PVY^N/NTN/N:O. This study supports previous evidence of a common origin for PVY^N/NTN in Europe and Brazil.     

Hemoplasmas in wild canids and felids in Brazil

Hemotropic mycoplasmas, epicellular erythrocytic bacterial parasites lacking a cell wall, are the causative agents of infectious anemia in numerous mammalian species. The presence of hemotropic mycoplasmas in blood samples of neotropical and exotic wild canids and felids from Brazilian zoos were recorded using molecular techniques. Blood samples were collected from 146 Brazilian wild felids, 19 exotic felids, 3 European wolves (Canis lupus), and from 97 Brazilian wild canids from zoos in the Brazilian states of S?o Paulo and Mato Grosso and the Federal District. Using conventional polymerase chain reaction (PCR), this work found 22 (13%) wild felids positive to Candidatus Mycoplasma haemominutum [4 jaguars (Panthera onca); 3 pumas (Puma concolor); 10 ocelots (Leopardus pardalis); 2 jaguarondis (Puma yagouaroundi); and 3 little spotted cats (Leopardus tigrinus)]. Only one little spotted cat (Leopardus tigrinus) was positive to Mycoplasma haemofelis, and none was positive to Candidatus Mycoplasma turicensis. Two bush dogs (Speothos venaticus) were positive for a Mycoplasma sp. closely related to Candidatus Mycoplasma haematoparvum, and two European wolves were positive for a Mycoplasma sp. closely related to Candidatus Mycoplasma haemominutum. This is the first study regarding the molecular detection of hemotropic mycoplasmas in wild canids.     

Pigeons (Columba livia) are a suitable experimental model for Neospora caninum infection in birds

Neospora caninum infections in chickens have been recently described by epidemiological and experimental approaches, and these birds may be considered natural intermediate hosts of the parasite. It has been postulated that other bird species might perform this role in wildlife as well. To better understand the sylvatic life cycle of N. caninum, further studies are required. In that sense, this work aimed to observe infection kinetics in pigeons experimentally infected with N. caninum. Experimental infections were conducted in parallel with a related protozoan, Toxoplasma gondii, which has been already described as able to infect pigeons in nature. Our results demonstrated that N. caninum disseminated through various tissues of this host and induced parasite-specific IgG seroconversion. Infection parameters were similar to that observed in the T. gondii infected group, although N. caninum-infected pigeons presented lower IgG titers during acute phase. The results herein described demonstrate that pigeons are a suitable model for N. caninum infection, considering that these data are in agreement with those observed in chickens experimentally infected with this parasite. As pigeons may be revealed as important reservoirs for N. caninum infection in nature, future studies are necessary to determine the real prevalence of this parasite in this and other birds in wildlife.     

Antioxidant in Mango (<Emphasis Type="Italic">Mangifera indica</Emphasis> L.) Pulp

This work was carried out to investigate the pulp composition of four mango cultivars (Haden, Tommy Atkins and Ubá) at the ripening stage in relation to three components with antioxidant potential (total phenolics, carotenoids and ascorbic acid). Total phenolic compound content was estimated by the Folin-Ciocalteu reagent and total carotenoid content by spectrophotometry at 450 nm. The contents of β-carotene and total vitamin C (ascorbic acid and dehydroascorbic acid) were quantified by high performance liquid chromatography. Differences were found among the four mango cultivars in all the components analyzed. The content of phenolic compounds ranged from 48.40 (Haden) to 208.70 mg/100 g (Ubá); total carotenoid from 1.91 (Haden) to 2.63 mg/100 g (Palmer); β-carotene from 661.27 (Palmer) to 2,220 μg/100 g (Ubá) and total ascorbic acid ranged from 9.79 (Tommy Atkins) to 77.71 mg/100 g (Ubá). These results corroborated previous information that mangoes are a good source of antioxidants in human diet.     

Study of lobation and vascularization of the lungs of wild boar (Sus scrofa)

The purpose of this study was to describe the anatomy of the lungs of wild boars for comparison with those of domestic swine. It was found that the right lung of the wild boar is divided into four lobes: cranial, median, caudal and accessory, whereas the left lung is divided into two lobes: cranial and caudal. In 93.4% of the cases, right pulmonary artery separates into the ascendant, descendant, median, accessory and caudal branches. In 73.3% of the cases, left pulmonary artery separates most frequently to form three branches to the cranial lobe, whereas the median lobe is generally supplied by only one arterial branch. There is a single pattern of bronchial distribution: in the right lung a tracheal bronchus leads to the cranial lobe, where it separates into the cranial and caudal bronchi and there are also bronchi to the median, caudal and accessory lobes. In the left lung, the large bronchus separates to form two branches, one of which further separates to form two branches to the cranial lobe whereas the other forms a single branch to the caudal lobe.     

Visceral leishmaniasis in a captive crab-eating fox Cerdocyon thous

Visceral leishmaniasis (VL) is a zoonotic disease with worldwide distribution. The crab-eating fox (Cerdocyon thous) is considered a wild reservoir of many zoonotical diseases, particularly VL. This study reported the presence of Leishmania infantum amastigotes in different organs of one captive C. thous found dead in a zoo. This animal was positive by the indirect fluorescence antibody test and had many clinical signs of VL. Intracellular amastigote forms of L. infantum were seen in neutrophils and macrophages in sample tissues from skin, lymph nodes (popliteal, submandibular, prescapular, and mesenteric), spleen, and liver. The numbers of positive cells and intracellular parasites were higher in macrophages than in neutrophils. In addition, polymerase chain reaction demonstrated extensive distribution of Leishmania DNA in C. thous tissues from multiple organs. The presence of intracellular amastigotes in neutrophils and macrophages as well as DNA of the parasite in tissues, specifically skin demonstrate that this crab-eating fox is an adequate host for L. infantum and reinforce the importance of VL for symptomatic wild canids kept in captivity in endemic areas.     

Survey for natural Neospora caninum infection in wild and captive birds

Neospora caninum is a protozoan parasite that presents worldwide distribution and is mainly implicated as responsible for bovine abortion. Although the presence of birds in cattle-raising properties is positively correlated to higher infection rates, very little has been described about the role of these animals in the parasite's life cycle. In that sense, this work aimed to investigate the serological and histological positivity of different avian species sampled in its natural habitat or in captivity. No serological positivity was observed in the 294 tested serum samples. On the other hand, Apicomplexa-like cysts found in muscular tissues of two Psittaciformes were immunostained with N. caninum antisera. These findings indicate that N. caninum may infect a wider range of hosts than described to date, and that further studies should be performed in order to determine the presence of the infection in different avian species.     

A morphological classification proposal for curimba (Prochilodus lineatus) sperm damages after cryopreservation

The present study evaluated some cryoprotectants and concentrations for curimba (Prochilodus lineatus) semen preservation from a simple methodology of analysis of sperm morphology. Semen of nine males was diluted (1:4) into eight cryosolutions, all of them containing 5% of BTS^? Minitüb, still comprising methanol or dimethyl sulphoxide (DMSO) at four concentrations: 5%, 7.5%, 10% and 12.5%. Morphological sperm analysis was performed on an optical microscope at × 1000 magnification. Damages on the head, midpiece and tail rest (the main and terminal portions) were investigated and classified as primary or secondary, according to the assumed influence on fertilization. There was a linear reduction in the total damages with an increase in the cryoprotectant concentration. Samples with DMSO showed the lowest percentage of fractured tails and tail stumps. The cryopreservation protocol retained the fertilizing potential of the sperm after freezing and both methanol 5% and DMSO 7.5% conferred adequate protection to the curimba sperm cells. Sperm morphology, as pointed out here, must be incorporated into routine fish sperm (cryopreserved or not) analysis once sperm defects influence fertilization directly. To the best of our knowledge, this was the first report on curimba sperm damages after cryopreservation by optical microscopy. The classification model presented in this study can be adequate to tackle fish sperm damages.