Ripening-related defense proteins in Annona fruit

In order to obtain a better understanding of the active defense strategy of cherimoya (Annona cherimola Mill.) fruit, hydrolytic and antifungal activity, as well as expression of proteins functionally and immunogenically related to the pathogenesis-related proteins chitinase (PR-Q) and 1,3-β-glucanase (PR-2), were estimated in fruit at different ripening stages. Increase in expression of the 27 kDa constitutive chitinase and the induction of two new proteins, a 26 kDa chitinase and a 51 kDa 1,3-β-glucanase were associated with enhanced in vitro hydrolytic and antifungal activity of the acidic protein extract in ripe fruit. Ripening modified the expression of constitutive basic isoenzymes, with a sharp decrease in both relative accumulation and hydrolytic activity. Likewise, a new basic 33 kDa chitinase was induced in the over-ripe fruit, concomitant with accumulation of a basic constitutive 76 kDa 1,3-β-glucanase. At this stage, the basic protein extract modified in vitro growth inhibition of Botrytis cinerea. Short-term high CO₂ treatment delayed fruit ripening and maintained a similar distribution of activity and isoenzymatic pattern in both protein fractions to that in unripe fruit. These results indicate that the changes in the pattern of defense proteins and hydrolytic activity in cherimoyas appear to be associated with ripening. Moreover, unlike the constitutively expressed isoenzymes, only the transitorily induced chitinases and 1,3-β-glucanases were associated with an active defense-related response.     

Detritivorous earthworms modify microbial community structure and accelerate plant residue decomposition

The impact of detritivorous earthworms (Eisenia andrei) on microbial community structure and function in grape marc, a lignocellulosic enriched plant residue, was investigated in a mesocosm experiment. Analysis of carbon and nitrogen pools was also carried out in order to evaluate how changes in microbial communities affect plant residue decomposition. The grape marc was completely processed after fifteen days as a result of the high density of earthworms present and the rapid gut transit time. Eisenia andrei had a large impact on the structure of the microbial community, as revealed by phospholipid fatty acid analysis. Earthworm activity reduced the abundance of both bacterial (except for Gram-negative bacteria) and fungal PLFA biomarkers relative to the control values. Decreases in microbial activity and in protease and cellulase activities were also attributable to the presence of earthworms. Moreover, earthworms strongly modified the substrate utilization patterns of microbial communities, as revealed by BIOLOG analysis. The presence of earthworms led to an increase in the utilization of some amino acids and polymers, which reached a higher substrate diversity value than that in the control mesocosm. The differences in microbial communities were accompanied by a reduction in the total C content and the labile C pool, relative to the control, although there were no significant differences in either cellulose or hemicellulose contents. However, total N content increased in both mesocosms – with and without earthworms – and the concentration of NH₄⁺ was also enhanced by earthworm activity. The results indicate that detritivorous earthworms play a key role in decomposing fresh plant residues in the short term via their intensive interactions with microbial communities.     

Chemical composition of the Lippia origanoides essential oils and their antigenotoxicity against bleomycin-induced DNA damage

The present work evaluated the chemical composition of the essential oils (EO) obtained from Lippia origanoides and their DNA protective effect against bleomycin-induced genotoxicity. L. origanoides EO chemical composition was determined by gas chromatography–mass spectrometry (GC–MS). The major compounds of the L. origanoides EOs were thymol (34–58%) and carvacrol (26%). The antigenotoxic effects of the EOs, major compounds and standard compound (epigallocatechin gallate) were assayed in co-incubation procedures using the SOS chromotest in Escherichia coli. Both EOs and their major compounds protected bacterial cells against bleomycin-induced genotoxicity indicating that these two compounds were principally responsible for the antigenotoxicity detected in the oils. Thymol and carvacrol antigenotoxicity was lower than those observed with epigallocatechin gallate. The results were discussed in relation to the chemopreventive potential of L. origanoides EOs and their major components, carvacrol and thymol.     

Bioprospection of Microalgae and Cyanobacteria as Biocontrol Agents Against Vibrio campbellii and Their Use in White Shrimp Litopenaeus vannamei Culture

Vibrio spp. are the most common and harmful shrimp pathogenic bacteria; however, microalgae and cyanobacteria have the ability to produce antimicrobial substances against these species. In this study, the organic and aqueous extracts of 28 species of marine microalgae and cyanobacteria were screened against Vibrio campbellii M1. Two of these phytoplankton species with antibacterial activity in aqueous extracts (Dunaliella tertiolecta and Skeletonema costatum) and nontoxic to brine shrimp Artemia franciscana nauplii were used to evaluate their anti‐Vibrio effect when used as green‐water cultures in Vibrio‐challenged white shrimp Litopenaeus vannamei cultures. No differences in mortality of juvenile L. vannamei were observed between treatments tested, suggesting that the pathogenicity of V. campbellii could be related to the growth stage of shrimp. The proximal composition of D. tertiolecta and S. costatum was in the recommended range for penaeid shrimp nutrition, allowing shrimp supplemented with these microalgae to have significantly greater total length and weight than control shrimp. Shrimp supplemented with S. costatum presented the highest values of organic mass (11.48 mg/organism) and growth rate (0.31 mg/d) in comparison to D. tertiolecta. These results indicate that microalgae are not only capable of producing antibacterial compounds against Vibrio but can also help shrimp nutrition.     

Progeny Production of the Copepods Pseudodiaptomus euryhalinus and Tisbe monozota in Monospecific and Mixed Cultures

The objective of this research was to culture the calanoid copepod Pseudodiaptomus euryhalinus, Johnson 1939, and the harpacticoid Tisbe monozota, Bowman, 1962, in monospecific and combined cultures (P. euryhalinus : T. monozota at 1:1, 2:1, and 1:2 starting ratios), and to compare the nauplii, copepodite, and adult production. Mean total production ranged from 740.3 ± 137.5 organisms/L (P. euryhalinus) to 884.3 ± 489.7 organisms/L (T. monozota) for monospecific cultures. The 1:1 ratio mixed cultures gave 780.4 ± 155.8 organisms/L, those with the 2:1 and 1:2 P. euryhalinus and T. monozota starting ratios produced 710.1 ± 195.2 and 799.7 ± 232.5 organisms/L, respectively, and there were no significant difference among treatments. All mixed cultures gave significantly lower copepodite productions than the monocultures of each species. In addition, the tendency to a decreased progeny production of the initial females of T. monozota indicates that the outcome of long‐term mixed cultures would production either a high dominance of P. euryhalinus, or monocultures of this species .     

对FC株猪源性肠毒素型大肠杆菌致病因子的研究

FC菌株是一株从腹泻仔猪粪便中分离的肠毒素型大肠杆菌(Enterotoxigenic E.coli,ETEC)。在MRHA反应中,本菌能凝集人O型、豚鼠、马、绵羊、牛、鸡和兔的红细胞,对人O型和豚鼠红细胞有很高的血凝性,血抗K88和K99血清不能抑制其对豚鼠和绵羊红细胞的血凝。在体外小肠上皮细胞吸附试验中,本菌对仔猪小肠上皮细胞具有强烈的吸附作用;透射电镜和扫描电镜观察证实了FC株菌除表面具有一种纤毛样结构外,还能定居在仔猪小肠段。血清学试验结果表明,本菌的O抗原属于O101。K88和987P两种抗血清均不能凝集本菌,而K99和F41抗血清均可凝集。对纯化的FC株菌粘着素抗原作等电聚焦和聚丙烯酰胺凝胶电泳分析,结果表明,该菌的粘着素是由等电点分别为4.61和9.78,分子量分别为29500和17500的两种蛋白质抗原所组成。此外,用乳鼠胃内投服试验和兔肠结扎试验证明,该菌只产生热稳定肠毒素。总之,本菌是一株能产生ST的K99,F41的肠毒素型大肠杆菌。     

Leishmaniasis in Canis familiaris – a Report About Microvascular and Cellular Architecture of the Infected Spleen

Leishmaniases are a globally widespread group of parasitic diseases from the group of anthropozoonoses. They are caused by an intracellular protozoan parasite that causes a wide spectrum of diseases in humans and dogs worldwide. In fact, the dog is considered one of the most important reservoir. The spleen is one of the several haematopoietic and immunocompetent organs involved. As this organ is a blood filter, the authors looked for the expression of this infection over the microvascular environment and modifications of the spleen cell population related to immunological responses to this parasitic condition.
Tools:  Scanning electronic microscopy over intact tissue and corrosion casts, transmission electronic microscopy, histology and immunohistochemistry (MAC 387 and CD3).
Results:  Our results show three important modifications concerning the microvascular architecture of the spleen when compared to the normal pattern. A striking scarcity of the sinusoidal system sheath that surrounds the central arteries of the white pulp, a huge development of pulp venules and veins, differentiation of typical features of high endothelial venular cells.
Remarks:  According to our results it seems that independent of the virulence of the parasite involved and the kind of immunity prevalent in a particular host, the spleen develops blood dynamic conditions that allows a sluggish blood flow so that cells involved in immunological processes can proliferate and differentiate and it also contributes to the trapping of lymphocytes to the area through the differentiation of typical characteristics of HEV endothelial cells.     

Use of touch-down polymerase chain reaction to enhance the sensitivity of Mycobacterium bovis detection.

The confirmatory diagnosis of Mycobacterium bovis (M. bovis) in animal samples is carried out by culture in Stonebrink media. However, culture is very slow because of the extremely long duplication time of the bacillus and difficult because of the scarcity of bacilli in diagnostic samples. This study describes the development of a single-tube touch-down polymerase chain reaction (PCR) protocol for the detection of M. bovis using primers that target the IS6110 element. Spiked water and milk as well as routine diagnostic samples (milk and nasal swabs) from M. bovis-positive cattle were tested. This protocol allows the rapid and sensitive detection of M. bovis in bovine samples by enhancing the sensitivity of standard PCR amplification.