Biometrical, biological, biochemical and molecular characteristics of Meloidogyne incognita isolates and related species

Meloidogyne incognita is one of the most polyphagous species of root-knot nematodes occurring in Brazil and worldwide. Eight M. incognita isolates were studied, representing two enzymatic phenotypes (esterase and malate desydrogenase: I1/N1, I2/N1) and four cryptic Meloidogyne sp.1 (S2/N1) isolates, representing one cytological type (3n?=?40–46). Three M. hispanica isolates (Hi3/N1, 2n?=?32–36) and two of an atypical Meloidogyne sp.2 (S2a/N3, 3n?=?40–44) were included in this study for comparison. All isolates were tested with three M. incognita-specific molecular markers. The primer pairs B06F/R, miF/R and incK14F/R amplified three species-specific fragments of 1,200?bp, 955?bp and 399?bp, respectively for M. incognita and Meloidogyne sp.1 isolates. No amplification occurred in the M. hispanica and Meloidogyne sp.2 isolates, except with primers miF/R (1,650?bp). The genetic variability of the Meloidogyne spp. isolates was evaluated, using RAPD and ISSR markers. The phylogenetic analyses revealed two strongly supported monophyletic clades: clade I, consisting of M. hispanica and the atypical Meloidogyne sp.2 isolates, and clade II, clustering together all M. incognita and the Meloidogyne sp.1 isolates. Considering the biometrical, cytological and molecular approaches, it was possible to conclude that the isolates with three enzymatic phenotypes (I1/N1, I2/N1 and S2/N1) presented the characteristics described for M. incognita. Some correlations were detected between the isozymatic phenotypes and the tree topology (S2a/N3, Hi3/N1, I1/N1, S2/N1), but no strict correlation could be observed for the phenotype I2/N1 and one isolate of S2/N1. Morphologically, the Msp.2 isolates differ from M. incognita and M. hispanica by the female stylet features presenting straight cone tip and round pear shaped knobs, posteriorly sloping. The results of this study suggested that the Msp.2 isolates with phenotypes S2aN3 belong to a new or an unidentified species closely related to M. hispanica.     

Assessment of the ideal ratios of digestible essential amino acids for pacu,Piaractus mesopotamicus,juveniles by the amino acid deletion method

The present study aimed to determine the ideal ratios of digestible essential amino acids (EAAs) for pacu (Piaractus mesopotamicus) juveniles by the amino acid (AA) deletion method. A completely randomized design which consisted of 11 treatments and three replicates each was used. The treatments included a control diet (CD) containing 55% of nonpurified natural ingredients and 45% of purified synthetic amino acids and ingredients, and other ten isonitrogenous and isoenergetic EAA limiting diets (LDs), each being deficient in 44.4 ± 0.02% of the respective EAA. Pacu juveniles with initial average body weight of 6.22 ± 0.09 g were distributed among 33 fiber glass tanks. Fish were fed with semipurified and extruded diets for 113 days two times a day until apparent satiation. The ideal ratio of each dietary EAA was calculated on the basis of the relationship between body N retention and amount of EAA deleted from the respective EAA LD. Based on the AA deletion method, the ideal ratios of digestible EAAs for pacu juveniles, relative to lysine requirement of 100% were estimated as: methionine 14.6%, threonine 35.0%, tryptophan 6.6%, arginine 62.8%, histidine 13.6%, isoleucine 26.3%, leucine 43.7%, phenylalanine 27.2%, and valine 35.8%.     

Detection of enzootic nasal tumor virus (ENTV) in a sheep flock in southern Brazil

Enzootic nasal tumor (ENT) is a contagious neoplasm associated with enzootic nasal tumor virus (ENTV), which may induce disease in sheep (ENTV-1) and goats (ENTV-2). This study aimed to describe the occurrence of ENT in two Texel sheep (Ovis aries) from a 75-sheep flock, located in the city of Gravataí, southern Brazil. Animals used to be purchased from different origins, and no specific tests for disease monitoring or quarantine procedure were performed. Affected animals presented respiratory distress, anorexia with severe weight loss, and mucopurulent unilateral nasal discharge. Necropsy was performed in both animals and nasal cavity masses were observed. Histopathological analysis demonstrated an epithelial neoplasm compatible with nasal adenocarcinoma. PCR using a protocol that amplifies a 591 bp sequence of 5’LTR-gag region of ENTV1 was performed followed by DNA sequencing. Both samples were positive, and the sequences obtained presented highest identity (97%) with ENTV strain TN28 (GenBank accession number MH899613) detected in a Texel sheep from Scotland. This is the first report of ENTV-1 leading to enzootic nasal tumor in sheep in Latin America, which confirms the presence of the retrovirus in sheep flocks in the Brazilian territory.

     

Ellagic Acid and Polyhydroxylated Urolithins Are Potent Catalytic Inhibitors of Human Topoisomerase II: An in Vitro Study

Ellagic acid (EA), a natural polyphenol abundant in fruits and common in our diet, is under intense investigation for its chemopreventive activity resulting from multiple effects. EA inhibits topoisomerase II, but the effects on the human enzyme of urolithins, its monolactone metabolites, are not known. Therefore, the action of several synthetic urolithins toward topoisomerases II was evaluated, showing that polyhydroxylated urolithins, EA, and EA-related compounds are potent inhibitors of the α and β isoforms of human topoisomerase II at submicromolar concentrations. Competition tests demonstrate a dose-dependent relationship between ATP and the inhibition of the enzyme. Docking experiments show that the active compounds bind the ATP pocket of the human enzyme, thus supporting the hypothesis that EA and polyhydroxylated urolithins act as ATP-competitive inhibitors of human topoisomerase II.     

Assessment of induction and recovery times of anaesthesia in Astyanax bimaculatus using 2‐phenoxyethanol and the essential oils of Melaleuca alternifolia and Ocimum gratissimum

This study aimed to determine the lowest effective concentration (LEC) of 2‐phenoxyethanol (2‐Phe) and essential oils (EOs) of Melaleuca alternifolia and Ocimum gratissimum able to induce anaesthesia in Astyanax bimaculatus. LEC was assumed to be the required concentration to carry out anaesthesia in less than 3 min with recovery in <5 min and sufficient to carry out routine aquaculture procedures. The following concentrations were tested: 2‐Phe (100, 200, 300, 400, 500 and 600 µl/L); M. alternifolia (200, 300, 400, 500 and 600 µl/L); and O. gratissimum (10, 20, 30, 40, 50, 60, 70, 100, 150, 250 and 300 µl/L). Results show that all three agents evaluated have effective anaesthetic properties for use in Astyanax bimaculatus with LEC for 2‐Phe, M. alternifolia and O. gratissimum of 200, 300 and 60 µl/L respectively.     

Estimating state-wide biomass carbon stocks for a REDD plan in Acre, Brazil

As in many other developing countries, the state government of Acre, Brazil, is developing a program for compensating forest holders (such as communities of rubber tappers and indigenous peoples as well as small, medium and large private land holders) reducing their emission of atmospheric heat-trapping gases by not deforesting. We describe and then apply to Acre a method for estimating carbon stocks by land cover type. We then compare the results of our simple method, which is based on vegetation mapping and ground-based samples, with other more technically demanding methods based on remote sensing. We estimated total biomass carbon stocks by multiplying the measured above-ground biomass of trees >10 cm DBH in each of 18 forest types and published estimates for non-forest areas, as determined by measurement of 44 plots throughout the state (ranging from 1 to 10 ha each), by land-cover area estimated using a geographical information system. State-wide, we estimated average above-ground biomass in forested areas to be 246 ± 90 Mg ha⁻¹; dense forest showed highest (322 ± 20 Mg ha⁻¹) and oligotrophic dwarf forest (campinarana) the lowest biomass (20 ± 30 Mg ha⁻¹). The two most widespread forest types in Acre, open canopy forests dominated by either palms and bamboo (for which ground-based data are scant), support an estimated 246 ± 44 and 224 ± 50 Mg ha⁻¹ of above-ground biomass, respectively. We calculate the total above-ground biomass of the 163,000 km² State of Acre to be 3.6 ± 0.8 Pg (non-forest biomass included). This estimate is very similar to two others generated using much more technologically demanding methods, but all three methods, regardless of sophistication, suffer from lack of field data.     

Real-time polymerase chain reaction based on msa2c gene for detection of Babesia bovis

This paper reports a quantitative real-time polymerase chain reaction (q-PCR) based on the msa2c gene and standardized with Platinum SYBR Green/ROX for the detection of Babesia bovis in cattle. The msa2c q-PCR amplified a DNA fragment with average dissociation temperature of 77.41°C (± 0.25°C). No amplification was detected when DNA from B. bigemina, A. marginale or Bos taurus was used as the template. The detection limit of the msa2c q-PCR was 1000 copies per ml of blood sample, with a linear correlation between the number of msa2c copies and threshold cycle. The comparison between msa2c q-PCR and conventional PCR for cytochrome b revealed 88.8% agreement, with a Kappa index of 0.75. In the comparison between msa2c q-PCR and an enzyme-linked immunosorbent assay (ELISA) with semi-purified B. bovis antigen, agreement was 96.3% and the Kappa index was 0.91. The agreement between three tests was 85.8%. The msa2c q-PCR detected a higher number of positive cattle than conventional PCR in an enzootically stable area, but did not differ significantly from ELISA. No significant differences were detected between the three diagnostic tests with cattle from an enzootically unstable area. All animals raised on a tick-free facility were negative for B. bovis in the three tests. These results suggest that msa2c q-PCR is a useful test for the detection of B. bovis infection.     

Using the same fish with different rules: A science‐based approach for improving management of recreational fisheries in a biodiversity hotspot of the Western South Atlantic

In this study we tested the hypothesis that Centropomus parallelus Poey moves between areas where fishing tourism is important, but the management rules for recreational fisheries are different. Fieldwork was conducted from May 2014 to June 2017 in an estuary located in the border between the states of São Paulo and Paraná, South‐Southeastern Brazil. Through a mark and recapture experiment, 2.000 individuals were tagged, of which 42 were recaptured. Among these, six fish were recaptured in Paraná, after being tagged in São Paulo, and a fish was recaptured in São Paulo, after being tagged in Paraná. These results support the hypothesis tested and indicate an uneven use of C. parallelus by anglers in both states. Thus, the use of the species and the recreational fisheries regulations need to be reviewed in the study area, taking into account the ability of C. parallelus to move across political boundaries and the importance of a fish‐oriented fisheries management regime.     

The role of açaí (Euterpe oleracea Mart. 1824) as a chemoprotective agent in the evaluation of antioxidant defence,oxidative damage and histology of juvenile shrimp Litopenaeus vannamei (BOONE, 1931) exposed to ammonia

This study investigated the effects of the use of the inclusion of açaí on the diet of shrimp Litopenaeus vannamei on antioxidant and histopathological responses after exposure to ammonia. The shrimps were fed two experimental diets: control and with 10% of açaí inclusion (W/W), for 35 days. Afterwards, the organisms were exposed at four concentrations of ammonia (0.01‐control; 0.26; 0.48 and 0.91 mg NH₃‐N L^?1) for 96 hr. The total antioxidant capacity (ACAP) of the gills decreased significantly in both diets when exposed to ammonia, whereas in the muscle, the açaí promoted an increase in ACAP. Concomitantly, lipid peroxidation levels increased in the gills and decreased in muscle. After exposure to ammonia, glutathione‐S‐transferase activity increased in hepatopancreas in shrimps fed with açaí facilitating the detoxification of lipid peroxidation by‐products, and the concentration of protein sulfhydryl groups decreased in the gills and muscle of the shrimp of the control diet, evidencing protein damage, an unobserved response in shrimps that received the açaí diet. Histopathological changes decreased in açaí‐fed shrimps about the control diet after exposure to ammonia. It is concluded that açaí mitigated ammonia‐induced histopathological changes, improved the antioxidant defence system (gills and muscle) and attenuated the lipid damage in the muscle.