Preliminary Study on the Activity of Phycobiliproteins against Botrytis cinerea

Phycobiliproteins (PBPs) are proteins of cyanobacteria and some algae such as rhodophytes. They have antimicrobial, antiviral, antitumor, antioxidative, and anti-inflammatory activity at the human level, but there is a lack of knowledge on their antifungal activity against plant pathogens. We studied the activity of PBPs extracted from Arthrospira platensis and Hydropuntia cornea against Botrytis cinerea, one of the most important worldwide plant-pathogenic fungi. PBPs were characterized by using FT-IR and FT-Raman in order to investigate their structures. Their spectra differed in the relative composition in the amide bands, which were particularly strong in A. platensis. PBP activity was tested on tomato fruits against gray mold disease, fungal growth, and spore germination at different concentrations (0.3, 0.6, 1.2, 2.4, and 4.8 mg/mL). Both PBPs reduced fruit gray mold disease. A linear dose–response relationship was observed for both PBPs against disease incidence and H. cornea against disease severity. Pathogen mycelial growth and spore germination were reduced significantly by both PBPs. In conclusion, PBPs have the potential for being also considered as natural compounds for the control of fungal plant pathogens in sustainable agriculture.     

Genetic analysis of Aequipecten opercularis and Mimachlamys varia (Bivalvia: Pectinidae) in several Atlantic and Mediterranean localities, revealed by mitochondrial PCR-RFLPs: a preliminary study

Aequipecten opercularis (Queen scallop) and Mimachlamys (Chlamys) varia (Black scallop) are important natural resources occurring in Atlantic and Mediterranean coasts. To develop an optimal sustainable exploitation plan, it is important to study the genetic structure of the different populations. In this study, we used polymerase chain reaction‐restriction fragment length polymorphisms for the determination of the genetic variation and population structure of these two species in different localities. Ten composite haplotypes were generated for A. opercularis and 15 haplotypes for M. varia. Of these, six and four were unique respectively. The analysis of the distribution of the different haplotypes between the localities showed no clear evidence of subdivision in A. opercularis, while in M. varia the results indicated that the two localities analysed should be managed as separate stocks.     

Effects of formalin on total haemocytes count and histopathological changes in the shrimp Litopenaeus schmitti (Pérez‐Farfante & Kensley 1997)

Formalin is used in shrimp industry to eliminate ectoparasites and as a disinfectant. However, the effect of formalin on shrimp's defense mechanism is unknown. Several biomarkers were used to assess the immune response of juveniles of the shrimp Litopenaeus schmitti exposed to different concentrations of formalin (10, 25 and 50 mg L^?1) during 24 and 48 h. Formalin concentrations of 25 and 50 mg L^?1 produced a significant reduction in the total haemocyte count at both 24 and 48 h. Peroxidase and phenoloxidase activity did not show variations. The total protein concentration in haemolymph of shrimps was not significantly different between experimental groups. Histological analysis showed hyperplasia in the gills of animals exposed at 10 mg L^?1 for 24 h; however, the exposure at 25 and 50 mg L^?1 caused necrosis in gills, hepathopancreas and the antennal gland.     

Streptomyces sp., a cause of fistulous withers in donkeys

Four of 10 donkeys, which showed lesions simulating fistulous withers, were examined clinically with the aim to cultivate and identify the causal agent. Aspiated purulent materials were subjected to bacteriological examination. The causal organisms were recovered in Tryptic Soya agar medium when incubated aerobically at 37 degrees C for up to 5 days. These organisms were found to be actinomycetes-like, Gam positive with stable branching filaments and to form heavy aerial hyphae on colony surface. The isolated organisms ere tentatively identified as Streptomyces sp. on the basis of morphological and cultural characteristics. The initial sequences analysis of the 16S rDNA gene conformed that one of the isolates (SD551) falls within the phylogenetic clade, which encompasses the genus Streptomyces. Studies are underway to further describe the disease and its causal agent. The report represents a good evidence to incriminate Streptomyces in the aetiology of the fistulous withers.     

Field and laboratory selection of Frankliniella occidentalis (Pergande) for resistance to insecticides

Response of western flower thrips, Frankliniella occidentalis (Pergande), to selection for resistance to insecticides commonly used to control this pest in Murcia (south-east Spain) was studied under field and laboratory conditions. In the field, plots within sweet pepper crops in commercial and experimental greenhouses were treated under different selection strategies: insecticide rotation versus formetanate reiteration, formetanate reiteration versus acrinathrin reiteration, and formetanate reiteration versus methiocarb reiteration. Thrips populations were sampled monthly and bioassayed against methiocarb, methamidophos, acrinathrin, endosulfan, deltamethrin and formetanate. In the laboratory, F occidentalis strains were selected against each insecticide for several generations. To evaluate cross-resistance, each selected strain was bioassayed with the other insecticides. Frankliniella occidentalis populations showed a rapid development of acrinathrin resistance, reaching high levels in field and laboratory conditions. Formetanate and methiocarb resistance were also observed, although development was slower and at moderate levels. Cross-resistances between acrinathrin/deltamethrin and acrinathrin/formetanate were detected under field and laboratory conditions. Formetanate/methiocarb cross-resistance was suspected in laboratory selections, but not in field assays. Simultaneous moderate resistance levels to the three specific insecticides against thrips (formetanate, methiocarb and acrinathrin) were shown in laboratory selection strains, indicating a general mechanism of resistance, probably metabolic.     

A PROPOSED RADIOGRAPHIC CLASSIFICATION SCHEME FOR CONGENITAL THORACIC VERTEBRAL MALFORMATIONS IN BRACHYCEPHALIC “SCREW‐TAILED” DOG BREEDS

Congenital vertebral malformations are common in brachycephalic “screw‐tailed” dog breeds such as French bulldogs, English bulldogs, Boston terriers, and pugs. The aim of this retrospective study was to determine whether a radiographic classification scheme developed for use in humans would be feasible for use in these dog breeds. Inclusion criteria were hospital admission between September 2009 and April 2013, neurologic examination findings available, diagnostic quality lateral and ventro‐dorsal digital radiographs of the thoracic vertebral column, and at least one congenital vertebral malformation. Radiographs were retrieved and interpreted by two observers who were unaware of neurologic status. Vertebral malformations were classified based on a classification scheme modified from a previous human study and a consensus of both observers. Twenty‐eight dogs met inclusion criteria (12 with neurologic deficits, 16 with no neurologic deficits). Congenital vertebral malformations affected 85/362 (23.5%) of thoracic vertebrae. Vertebral body formation defects were the most common (butterfly vertebrae 6.6%, ventral wedge‐shaped vertebrae 5.5%, dorsal hemivertebrae 0.8%, and dorso‐lateral hemivertebrae 0.5%). No lateral hemivertebrae or lateral wedge‐shaped vertebrae were identified. The T7 vertebra was the most commonly affected (11/28 dogs), followed by T8 (8/28 dogs) and T12 (8/28 dogs). The number and type of vertebral malformations differed between groups (P = 0.01). Based on MRI, dorsal, and dorso‐lateral hemivertebrae were the cause of spinal cord compression in 5/12 (41.6%) of dogs with neurologic deficits. Findings indicated that a modified human radiographic classification system of vertebral malformations is feasible for use in future studies of brachycephalic “screw‐tailed” dogs.     

Sorption of pesticides in tropical and temperate soils from Australia and the Philippines

The sorption behavior of diuron, imidacloprid, and thiacloprid was investigated using 22 soils collected in triplicate from temperate environments in Australia and tropical environments in Australia and the Philippines. Within the temperate environment in Australia, the soils were selected from a range of land uses. The average KOC values (L/kg) for imidacloprid were 326, 322, and 336; for thiacloprid, the values were 915, 743, and 842; and for diuron, the values were 579, 536, and 618 for the Ord (tropical), Mt. Lofty (temperate), and Philippines (tropical) soils, respectively. For all soils, the sorption coefficients decreased in the following order: thiacloprid > diuron > imidacloprid. There were no significant differences in sorption behavior between the tropical soils from the Philippines and the temperate soils from Australia. Sorption was also not significantly related with soil characteristics, namely, organic carbon (OC) content, clay content, and pH, for any of the three chemicals studied. When the data were sorted into separate land uses, the sorption of all three chemicals was highly correlated (P < 0.001) with OC for the rice soils from the Philippines. Sorption coefficients for all three chemicals were highly correlated with OC in temperate, native soils only when one extreme value was removed. The relationships between sorption of all three chemicals and OC in temperate, pasture soils were best described by a polynomial. Sorption coefficients for imidacloprid and thiacloprid determined in the temperate pasture soils remained fairly consistent as the OC content increased from 3.3 to 5.3%, indicating that, although the total OC in the pasture soils was increasing, the component of OC involved with sorption of these two compounds may have been remaining constant. This study demonstrated that the origin of the soils (i.e., temperate vs tropical) had no significant effect on the sorption behavior, but in some cases, land use significantly affected the sorption behavior of the three pesticides studied. The impact of land use on the nature of soil OC will be further investigated by NMR analysis.     

Virulence factors in Escherichia coli strains isolated from pigs in the Ribeirao Preto region, State of Sao Paulo, Brazil.

Three-hundred faecal swabs were obtained from pigs with diarrhoea in farms located in different areas of the Ribeirao Preto region in the State of Sao Paulo. One-hundred Escherichia coli strains were isolated and tested for production of thermolabile (TL) and thermostable (STRa and STb) enterotoxins, and for the presence of colonization factors F4, F5 and F6. The strains were also tested for sensitivity to 14 antibiotics and chemotherapeutic agents. Twenty-four Escherichia coli strains produced enterotoxin STb, 5 produced LT and 3 produced STa. In the mannose-resistant haemagglutination reaction, one strain reacted positively with sheep, chicken, horse and human red blood cells and another reacted positively with guinea pig, sheep, chicken, horse and human red cells. However, both strains were negative for colonization factors F4, F5 and F6 when submitted to the slide agglutination test. All Escherichia coli strains were resistant to at least one antibiotic, the highest percentages being obtained for resistance to penicillin, tetracycline and cephalotin. In addition to the importance of the virulence factors normally encountered in enterotoxigenic Escherichia coli strains from pigs, the present results show the possible existence of new colonization factors other than F4, F5 and F6 participating in E. coli-induced pigs colibacillosis in the Ribeirao Preto region.