1-O-Alkylglycerol Ethers from the Marine Sponge Guitarra abbotti and Their Cytotoxic Activity

The cytotoxicity-bioassay-guided fractionation of the ethanol extract from the marine sponge Guitarra abbotti, whose 1-O-alkyl-sn-glycerol ethers (AGEs) have not been investigated so far, led to the isolation of a complex lipid fraction containing, along with previously known compounds, six new lipids of the AGE type. The composition of the AGE fraction as well as the structures of 6 new and 22 previously known compounds were established using ¹H and ¹³C NMR, GC/MS, and chemical conversion methods. The new AGEs were identified as: 1-O-(Z-docos-15-enyl)-sn-glycerol (1), 1-O-(Z-docos-17-enyl)-sn-glycerol (2), 1-O-(Z-tricos-15-enyl)-sn-glycerol (3), 1-O-(Z-tricos-16-enyl)-sn-glycerol (4), 1-O-(Z-tricos-17-enyl)-sn-glycerol (5), and 1-O-(Z-tetracos-15-enyl)-sn-glycerol (6). The isolated AGEs show weak cytotoxic activity in THP-1, HL-60, HeLa, DLD-1, SNU C4, SK-MEL-28, and MDA-MB-231 human cancer cells. A further cytotoxicity analysis in JB6 P⁺ Cl41 cells bearing mutated MAP kinase genes revealed that ERK2 and JNK1 play a cytoprotective role in the cellular response to the AGE-induced cytotoxic effects.     

The importance of manual white blood cell differential counts and platelet estimates in elephant hematology: blood film review is essential

Unique features of elephant hematology are known challenges in analytical methodology like two types of monocytes typical for members of the Order Afrotheria and platelet counts of the comparatively small elephant platelet. To investigate WBC differential and platelet data generated by an impedance-based hematology analyzer without availability of validated species-specific software for recognition of elephant WBCs and platelets, compared to manual blood film review. Blood samples preserved in ethylenediaminetetraacetic acid (EDTA) of 50 elephants (n = 35 Elephas maximus and n = 15 Loxodonta africana) were used. A Mann-Whitney test for independent samples was used to compare parameters between methods and agreement was tested using Bland-Altman bias plots. All hematological variables, including absolute numbers of heterophils, lymphocytes, monocytes, eosinophils, basophils, and platelets, were significantly different (p < 0.0001) between both methods of analysis, and there was no agreement using Bland-Altman bias plots. Manual review consistently produced higher heterophil and monocyte counts as well as platelet estimates, while the automated analyzer produced higher lymphocyte, eosinophil, and basophil counts. The hematology analyzer did not properly differentiate elephant lymphocytes and monocytes, and did not accurately count elephant platelets. These findings emphasize the importance of manual blood film review as part of elephant complete blood counts in both clinical and research settings and as a basis for the development of hematological reference intervals.     

Virulence genes and plasmid profiles in Rhodococcus equi isolates from domestic pigs and wild boars (Sus scrofa) in Brazil

The virulence genes and plasmid profiles of 23 Rhodococcus equi isolates from 258 lymph nodes from domestic pigs (129 nodes with lesions and 129 without lesions) and 120 lymph nodes from slaughtered wild boars (60 nodes with lesions and 60 without) were characterized. R. equi was obtained from 19 lymph nodes of domestic pigs, 17 with, and two without lesions, and from four lymph nodes with lesions, from wild boars. The 23 isolates were tested for the presence of vapA and vapB genes, responsible for the 15–17 and 20 kDa virulence-associated proteins, respectively, by PCR in order to characterize as virulent (VapA), intermediately virulent (VapB) and avirulent. Plasmid DNAs were isolated and analyzed by digestion with restriction endonucleases to estimate size and compare their polymorphisms. Of the 19 domestic pigs strains, seven (36.8%) were avirulent and 12 (63.2%) were intermediately virulent, with the intermediately virulent isolates being plasmid types 8 (8 isolates), 10 (2 isolates), 1 (1 isolate) and 29 (1 isolate). The plasmid type of four strains isolated from wild boars was also intermediately virulent type 8. None of the domestic pigs and wild boar isolates showed the vapA gene. These findings demonstrate a high occurrence of plasmid type 8 in isolates from pigs and wild boars, and the similarity of plasmid types in the domestic pigs, wild boars and human isolates in Brazil.     

Ecological evaluation of polymetallic soil quality: the applicability of culture-dependent methods of bacterial communities studying

Purpose

This work aimed to study the effect of long-term polymetallic contamination on the state and parameters of soil bacterial communities, including the abundance of different groups of culturable bacteria and the activity of nitrification.

Materials and methods

Monitoring plots were located in the dry lake and surrounding area, which had been formerly used for the discharge of industrial waste. The soils in the 16 plots were characterized by extremely high levels of heavy metal pollution. This study evaluated the main soil physicochemical properties by various methods, total metal contents by X-ray analysis, mobile metal content by atomic absorption spectrophotometry, the abundance of chosen groups of culturable bacteria by inoculation on solid media, and nitrification activity from ammonium and nitrite oxidation rates.

Results and discussion

High adaptation capacity of microbial communities to long-term pollution was revealed through marked lack of decrease in the abundance of some of the bacterial groups in soils with high contamination levels. Among the bacteria determined by the colony count method, copiotrophic and spore-forming bacteria were the least sensitive to contamination, and actinomycetes were the most sensitive. The high levels of soil pollution with heavy metals had pronounced adverse effects on nitrification activity. The decrease in activity was strongly correlated with pollutant concentrations. The oxidation of nitrite was shown to be more affected by pollution that the oxidation of ammonium.

Conclusions

Some groups and parameters of culturable microorganisms can be used for soil status estimation under pollution conditions even though they are only a small fraction of the microbial community. The most sensitive parameter was the nitrification rate, while the number of actinomycetes was found to be most promising parameter among the groups of bacteria determined by plate counts. The use of sensitive groups of culturable microorganisms for bioindication purposes is a method, which may provide a cheap and sufficiently reliable tool for large-scale soil monitoring studies.

     

Anti HSV-1 Activity of Halistanol Sulfate and Halistanol Sulfate C Isolated from Brazilian Marine Sponge Petromica citrina (Demospongiae)

The n-butanol fraction (BF) obtained from the crude extract of the marine sponge Petromica citrina, the halistanol-enriched fraction (TSH fraction), and the isolated compounds halistanol sulfate (1) and halistanol sulfate C (2), were evaluated for their inhibitory effects on the replication of the Herpes Simplex Virus type 1 (HSV-1, KOS strain) by the viral plaque number reduction assay. The TSH fraction was the most effective against HSV-1 replication (SI = 15.33), whereas compounds 1 (SI = 2.46) and 2 (SI = 1.95) were less active. The most active fraction and these compounds were also assayed to determine the viral multiplication step(s) upon which they act as well as their potential synergistic effects. The anti-HSV-1 activity detected was mediated by the inhibition of virus attachment and by the penetration into Vero cells, the virucidal effect on virus particles, and by the impairment in levels of ICP27 and gD proteins of HSV-1. In summary, these results suggest that the anti-HSV-1 activity of TSH fraction detected is possibly related to the synergic effects of compounds 1 and 2.     

When the ground cover brings guests: is <Emphasis Type="Italic">Anaphothrips obscurus</Emphasis> a friend or a foe for the biological control of <Emphasis Type="Italic">Tetranychus urticae</Emphasis> in clementines?

Biological control of Tetranychus urticae Koch (Acari: Tetranychidae), a key pest of clementines, can be improved in this crop with the establishment of a ground cover of Festuca arundinacea Schreber (Poaceae). This cover houses an abundant and diverse community of predatory Phytoseiidae mites including Euseius stipulatus (Athias-Henriot), Neoseiulus barkeri Hughes, Neoseiulus californicus (McGregor) and Phytoseiulus persimilis Athias-Henriot and a dense population of the grass thrips Anaphothrips obscurus Müller (Thysanoptera: Thripidae) throughout the year. The aim of this study was to determine whether the presence of this thrips species could be related to the improvement in the biological control of T. urticae. Therefore, the capacity of the abovementioned phytoseiids to feed and reproduce on A. obscurus and their feeding preferences when T. urticae and A. obscurus were simultaneously offered, were analyzed. The results show that E. stipulatus, N. barkeri and N. californicus have a type II functional response when offered A. obscurus nymphs, whereas P. persimilis barely feeds on this thrips species. Furthermore, N. barkeri and N. californicus can reproduce feeding only on thrips. Regarding prey preference, the Tetranychus spp.-specialist P. persimilis preferably preyed on T. urticae, the generalists N. barkeri and E. stipulatus preferred A. obscurus, and the selective predator of tetranychid mites N. californicus showed no preference. Therefore, we hypothesize that the enhanced biological control of T. urticae observed could be related to A. obscurus becoming an alternative prey for non-specialist phytoseiids, without altering the control exerted by the T. urticae-specialist P. persimilis and likely reducing intraguild predation.