Investigation of biofilm production and icaA and icaD genes in Staphylococcus aureus isolated from heifers and cows with mastitis

Biofilm formation and antimicrobial resistance of Staphylococcus aureus are important virulence factors in cases of mastitis in dairy cows. However, few studies have investigated mastitis strains isolated from heifers. Within this context, the objective of the present study was to investigate biofilm formation on Congo red agar, the presence of the icaA and icaD genes by polymerase chain reaction (PCR), and the percentage of in vitro antimicrobial resistance of 110 S. aureus isolates from mammary gland secretions of heifers and cows with mastitis. PCR detected the icaA and icaD genes in 98% and 100% of isolates, respectively. However, only 55.5% of all isolates produced a biofilm on Congo red agar. Antimicrobial susceptibility testing revealed that 47.0% of isolates from heifers and 70.4% of isolates from cows were resistant to at least one of the antimicrobial agents tested. Resistance to penicillin and/or ampicillin was the most frequent (44.5%). These results indicate the need to implement prophylactic and control measures of mastitis for heifers. Heifers and cows can carry resistant strains with the capacity of biofilm production, a fact representing a threat to public health and animal well‐being and generating losses to dairy farmers.     

Evaluation of hepatitis E virus infection between different production systems of pigs in Brazil

The objectives of this study were to (1) investigate the occurrence of hepatitis E virus (HEV) in pigs from large-scale and family-scale farms, (2) genetically characterize the strains isolated, and (3) study the pathogenesis of swine HEV infection via immunohistochemistry. A total of 50 pigs from 10 farms in Mato Grosso State, Brazil were divided according to type of production system into either large-scale farms (n?=?5) or family-scale farms (n?=?5). Samples of liver, gallbladder, small and large intestines, bile, and feces from the pigs were analyzed by nested PCR with primers targeting the ORF2 region of HEV and by immunohistochemistry. Of the eight HEV-positive samples from pigs of family-scale farms, phylogenetic analysis revealed that seven of the swine HEV isolates clustered with subtype 3b of genotype 3 and one isolate was categorized with subtype 3 f. The HEV antigen was detected mainly in the small intestine samples from family-scale farms, suggesting an early stage HEV infection. HEV was not detected in the samples of pigs from large-scale farms, reinforcing the need for additional studies to evaluate the risk of transmission of HEV to humans from pigs from family-scale farms in Mato Grosso State.     

The Use of Equine Follicle Stimulating Hormone to Increase Equine Chorionic Gonadotropin in the Pregnant Mare

Equine chorionic gonadotropin (eCG), obtained from pregnant mares, is used for assisted reproductive technologies in laboratory rodents and livestock. The objective of the present study was to use equine follicle-stimulating hormone (eFSH) to increase the incidence of twin pregnancies, through multiple ovulations, and increase eCG. Nineteen light horse–type mares were enrolled in the study. The control group (n = 9) was bred with fresh or cooled semen and given human chorionic gonadotropin (hCG) at the time of breeding. The second group (n = 10) was given 12.5 mg of eFSH intramuscularly twice a day beginning 5–7 days after ovulation. Prostaglandin F2α was administered intramuscularly the second day of eFSH treatment. Treatment with eFSH continued until follicles were >35 mm in diameter, and mares were then given no treatment for 36 hours. The mares were then bred with fresh or cooled semen from the same stallion as the control group and given hCG. Blood samples were taken weekly from day 35 to day 105 after ovulation. Serum concentration of eCG was obtained, and data were analyzed with multivariate analysis using the mixed procedure. Significance was set at P < .05. Data were combined for all mares carrying twins and compared with those carrying singletons. The group of mares carrying twins had higher peak concentrations of eCG and higher values for area under the curve compared with mares carrying singletons (P < .05). These results suggest inducing twins could be a method used to increase eCG production.     

Escherichia coli challenge and one type of smectite alter intestinal barrier of pigs

An experiment was conducted to determine how an E. coil challenge and dietary clays affect the intestinal barrier of pigs. Two groups of 32 pigs （initial BW： 6.9 ± 1.0 kg） were distributed in a 2 x 4 factorial arrangement of a randomized complete block design （2 challenge treatments： sham or E. coil, and 4 dietary treatments： control, 0.3% smectite A, 0.3% smectite B and 0.3% zeolite）, with 8 replicates total. Diarrhea score, growth performance, goblet cell size and number, bacterial translocation from intestinal lumen to lymph nodes, intestinal morphology, and relative amounts of sulfo and sialo mucins were measured. The E. coli challenge reduced performance, increased goblet cell size and number in the ileum, increased bacterial translocation from the intestinal lumen to the lymph nodes, and increased ileal crypt depth. One of the clays （smectite A） tended to increase goblet cell size in ileum, which may indicate enhanced protection. In conclusion, E. coli infection degrades intestinal barrier integrity but smectite A may enhance it.     

Genetic differentiation among Atlantic island populations of the brown spiny lobster Panulirus echinatus (Decapoda: Palinuridae)

1. Declines in Panulirus echinatus Smith, 1869 populations along their wide distribution in the Atlantic Ocean have spurred efforts to improve their fisheries management and conservation. In this study, the genetic structure of these populations is reported for the first time.
2. In a survey of 18 species‐specific polymorphic microsatellite markers, 152 individuals were genotyped from five Atlantic oceanic islands, covering most of the insular distribution range of the species. The analyses revealed that P. echinatus is genetically partitioned into two stocks in the Atlantic Ocean. A highly significant genetic structure was observed between north‐east and south‐west Atlantic populations based on fixation index, discriminant analysis of principal components, and structure and barrier analysis.
3. We suggest that the Equatorial Circulation System represents a biophysical barrier that effectively limits migration among Atlantic subtropical gyre systems, as has been described for other species. Other physical and ecological barriers, such as the Mid‐Atlantic Ridge itself, the distance between the eastern and western sides of the Atlantic (Mid‐Atlantic Barrier) and water mass differences, as well as other biological aspects, may also influence larval dispersal and modulate the insular distribution of this species.
4. The results show the existence of two distinct genetic stocks of P. echinatus and have implications for fisheries management in the Atlantic Ocean, including their independent management according to their individual status. The Cabo Verde and the Canarian populations (north‐east Atlantic) showed the lowest level of genetic variability in comparison with the south‐western populations. A combination of factors that have occurred or are occurring in the Canary Islands, such as overfishing and volcanic eruptions, is likely to explain the reduced abundance of this lobster species in the area.

     

Herbivory, pathogens, and epiphylls in Araucaria Forest and ecologically-managed tree monocultures

Over the last centuries, natural forests have been replaced by extensive and homogeneous tree monocultures that cause strong impacts on ecological interactions and ecosystem processes. We expect, however, that ecologically-sustainable management practices can help to mitigate these effects. This study investigates how the replacement of Araucaria Forest by ecologically-managed tree monocultures affects leaf damage patterns produced by external chewers, internal chewers, leaf miners, galling insects, and pathogens, as well as the incidence of epiphylls. The study was performed in the São Francisco de Paula National Forest (southern Brazil) on 12 1-ha plots of Araucaria Forest and ecologically managed plantations of Araucaria, Pinus and Eucalyptus. Overall, 9955 leaves of the understory community (up to 1 m tall) were sampled and analyzed. The mean and the frequency distribution pattern of leaf damage were very similar between Araucaria Forest and all three tree monocultures. Also, there was no difference between habitats in the percentage of leaves attacked by different feeding guilds of insect herbivores and by pathogens. Araucaria Forest had a higher percentage of leaves with epiphylls than Eucalyptus plantation, probably due to its shadier and moister microclimate. Ecologically managed tree monocultures sustain a wide variety of herbivorous insects, pathogens and epiphylls which helps the maintenance of key ecological interactions and the functioning of the ecosystem.     

Chemoenzymatic synthesis of homogeneous ultralow molecular weight heparins

Ultralow molecular weight (ULMW) heparins are sulfated glycans that are clinically used to treat thrombotic disorders. ULMW heparins range from 1500 to 3000 daltons, corresponding from 5 to 10 saccharide units. The commercial drug Arixtra (fondaparinux sodium) is a structurally homogeneous ULMW heparin pentasaccharide that is synthesized through a lengthy chemical process. Here, we report 10- and 12-step chemoenzymatic syntheses of two structurally homogeneous ULMW heparins (MW = 1778.5 and 1816.5) in 45 and 37% overall yield, respectively, starting from a simple disaccharide. These ULMW heparins display excellent in vitro anticoagulant activity and comparable pharmacokinetic properties to Arixtra, as demonstrated in a rabbit model. The chemoenzymatic approach is scalable and shows promise for a more efficient route to synthesize this important class of medicinal agent.     

Redistribution of 10B Absorbed by Leaves and Roots of Cashew Seedlings

Boron (B) is transported mainly in the xylem, but by using isotopic tracers it was possible to verify that B has significant mobility in the phloem of polyols-producing species. Thus, the objective of this study was to evaluate the mobility of ¹⁰B applied to cashew plants. The experiment was conducted from 05/10 to 07/10/2010 in a greenhouse of University of São Paulo State-UNESP, Ilha Solteira Campus, State of São Paulo, Brazil. 0.5 mg of B L^?1 of soil was used in the substrate. A solution of 255 mg of B L^?1 was used in recently mature and fully developed leaves. The total content of B and and percentage of boron in the plant derived from fertilizer (Bppf%) were evaluated in the old and new leaves stemming at 30 and 60 days after the application of ¹⁰B. As evidenced, B applied to leaves had restricted mobility in cashew tree. However, B applied in the substrate was absorbed and translocated to young leaves.     

Evaluation of antiviral activity of phenolic compounds and derivatives against rabies virus

Human rabies is a viral disease with a great impact on public health, mainly on account of its fatal course in the majority of cases. Despite the well-established prophylaxis by immunization, rabies is believed to be responsible for 40,000-70,000 human deaths per year, mostly in endemic areas. Palliative support and experimental protocols to avoid death have been employed with no expressive results, with the exception of a recent human case of recovery from rabies. No antiviral drugs are currently available to fight against this infection. In combination with the prophylaxis, an antiviral drug would be useful for human rabies treatment, providing enhanced protection against the encephalitis caused by the virus. Phenolic compounds are derived from the secondary plant metabolism, although they can also be obtained by synthetic processes. Many studies have shown a great range of pharmacological effects for these substances, including vasodilatation, antiallergenic, antiinflammatory and antiviral properties, among others. In this study, the potential in-vitro anti-rabies activity of 24 synthetic phenolic compounds was evaluated using McCoy cells and PV rabies strain. The cytotoxicity (CC50) was assayed by the MTT method and the antiviral activity (IC50) was estimated by the inhibition of viral cytopathic effects. Isoprinosine and ketamine were used as positive controls. The tested compounds showed selectivity indices (SI=CC50/IC50) ranging from 1.0 to 3.9. Six phenolic compounds failed to inhibit the cytopathic effect to any degree, and four showed SI > or = 3.0. According to these results, some probable structure-activity relationships are suggested. It was observed that the presence of free hydroxyl and ether groups influenced the anti-rabies activity. However, additional studies are required to establish these relationships.