Anisakis spp. in European hake, Merluccius merluccius (L.) from the Atlantic off north-west Africa and the Mediterranean off southern Spain

We studied the parasitization by Anisakis spp. in European hake (Merluccius merluccius) of 40–52 cm from the Atlantic off north-west Africa and the Mediterranean off southern Spain. Infection parameters differed: The fish from the Atlantic showing a prevalence of 87.97% and a mean intensity of 4.69, while, in those from the Mediterranean, these were 41.27% and 1.73, respectively. In both samples the two-third larval stage types were isolated: Anisakis simplex sensu lato and Anisakis larvae type II, with prevalence of 85.71% and 30.83% in fish from the Atlantic and 41.27% and 1.59% for those from the Mediterranean, respectively. In both samples, the prevalence of larvae in viscera was clearly higher than in the muscle tissue. We also observed an increase in parasitization with increasing host length, those ≥ 46 cm having the highest prevalences (94.87% for those from the Atlantic and 58.33% for those from the Mediterranean; p<0.03). This paper is dedicated to the memory of Prof. Dr Ignacio Navarrete López-Cózar, DVM, recently deceased.     

Isolation, characterization and chromosome localization of repetitive DNA sequences in bananas (Musa spp.)

Partial genomic DNA libraries were constructed in Musa acuminata and M. balbisiana and screened for clones carrying repeated sequences, and sequences carrying rDNA. Isolated clones were characterized in terms of copy number, genomic distribution in M. acuminata and M. balbisiana, and sequence similarity to known DNA sequences. Ribosomal RNA genes have been the most abundant sequences recovered. FISH with probes for DNA clones Radka1 and Radka7, which carry different fragments of Musa 26S rDNA, and Radka14, for which no homology with known DNA sequences has been found, resulted in clear signals at secondary constrictions. Only one clone carrying 5S rDNA, named Radka2, has been recovered. All remaining DNA clones exhibited more or less pronounced clustering at centromeric regions. The study revealed small differences in genomic distribution of repetitive DNA sequences between M. acuminata and M. balbisiana, the only exception being the 5S rDNA where the two Musa clones under study differed in the number of sites. All repetitive sequences were more abundant in M. acuminata whose genome is about 12% larger than that of M. balbisiana. While, for some sequences, the differences in copy number between the species were relatively small, for some of them, e.g. Radka5, the difference was almost thirty-fold. These observations suggest that repetitive DNA sequences contribute to the difference in genome size between both species, albeit to different extents. Isolation and characterization of new repetitive DNA sequences improves the knowledge of long-range organization of chromosomes in Musa. This revised version was published online in July 2006 with corrections to the Cover Date.     

Physical organization of the 18S and 5S ribosomal RNA genes in the mitochondrial genome of rye (Secale cereale L.)

Summary The mitochondrial 18S and 5S ribosomal RNA (rRNA) genes of rye, plus a total of about 90 kilobase pairs of flanking DNA, have been cloned and maps of restriction enzyme cleavage sites have been constructed. Like their homologs from hexaploid wheat, the rye genes are closely linked and are part of a three-copy family of recombining repeats (the 18S/5S repeat). The rye repeat probably also contains a mitochondrial tRNA^fMet gene, which the wheat repeat is known to carry. However, despite the overall organizational similarity between the wheat and rye 18S/5S repeats in the immediate vicinity of their coding regions, extensive rearrangement of flanking sequences has taken place during evolutionary divergence of the two species. Our data provide additional support for an emerging picture of plant mitochondrial genomes as evolving much more rapidly in structure than in sequence.     

An ultrastructural study of the microtriches of adultProteocephalus tidswelli (Cestoda: Proteocephalidea)

Ultrastructural observations on adultProteocephalus tidswelli revealed a marked microtrichial polymorphism. Structural and dimensional variations of microtriches between different regions of the strobila and scolex, as well as within the same region, were observed. The authors suggest that microtriches are involved in a diversity of functions, and possible functional activities are discussed.Abbreviations A Apical organ - B Base of microthrix - BL Blade-like microthrix - BP Baseplate - C Cytoplasmic core - D Disc bodies - DC Distal cytoplasm - FL Filamentous microthrix - G Gravid segments - GBL Giant blade-like microthrix - GC Granular cytoplasm - GX Glycocalyx - IM Immature segments - M Matrix - MA Mature segments - MF Microfilaments - N Neck - PL Peg-like microthrix - PM Plasma membrane - R Rostellum - S Sucker - SH Shaft of microthrix - SL Spine-like microthrix - ST Strap-like structures - T Thick-walled tube     

Effect of triterpene acids of Eriobotrya japonica (Thunb.) Lindl. leaf on inflammatory cytokine and mediator induction from alveolar macrophages of chronic bronchitic rats

Objective: The study was to evaluate the effect of triterpene acids of Eriobotrya japonica (thunb.) lindl. leaf (TAL) on inflammatory cytokine and mediator expression in alveolar macrophages (AM) of chronic bronchitic (CB) rats. Methods: CB was induced by endotracheal instillation of lipopolysaccharide (LPS) followed by Bacillus Calmette Guerin (BCG) injection via the caudal vein one week later. Treatment groups received TAL at there different doses (50, 150, or 450 mg/kg daily i. g.), Ketotifen fumarate (5 mg/kg daily i. g.) or dexamethasone (1.2 mg/kg daily i. g.) for two weeks, 7 days after LPS injection. AM were then isolated and incubated for 24 h. IL-1, TNF-α and PGE2 levels in cultured supernatants were measured by thymocyte co-stimulating assay and radioimmunoassay. Immunocytochemistry staining and western-blot were used for intracellular location and activation of p65 subunit of NF-kB. LTB₄ level was analyzed by reverse-phase high performance liquid chromatography (RP-HPLC). Results: The levels of TNF-α, IL-1, NF-kB, PGE2 and LTB₄ expression in AM of TAL groups were significantly decreased compared to the CB group (P < 0.05 or P < 0.01), in a dose dependent manner. Conclusion: TAL inhibited NF-kB activation in AM from CB rats and led to down regulation of TNF-α, IL-1, PGE₂ and LTB₄ expression, which might be a mechanism for its anti-inflammatory effects in CB rats. Received 17 November 2005; returned for revision 19 March 2006; returned for final revision 14 July 2006; accepted by G. Wallace 30 August 2006     

A radioiodinated peptidyl diazomethane detects similar cysteine proteinases in amastigotes and promastigotes ofLeishmania (L.) mexicana andL. (L.) amazonensis

Cysteine proteinase activities were examined in lesion amastigotes as well as in stationary-phase promastigotes ofLeishmania (L.) mexicana andLeishmania (L.) amazonensis isolates. Enzyme detection in gelatin gels revealed that amastigotes of threeL. (L.) mexicana isolates (M379, IOC-0561, and IP) shared similar proteinases, including the multiple low-molecular-weight (25–35 kDa) cysteine proteinases. High cysteine proteinase activity was also observed inL. (L.) amazonensis amastigotes, but the banding profile was different in two of the isolates examined. Promastigotes displayed fewer low-molecular-weight proteinase bands, and these were much less intense as compared with those of lesion amastigotes. Independently of theLeishmania isolates and developmental stages examined, incubation of the parasites for 2 h with 0.2 M radioiodinatedN-benzyloxycarbonyl-tyrosyl-alanyl diazomethane (Z-Tyr[¹²⁵I]-AlaCHN₂) markedly and selectively labeled bands comigrating with the 28- and 31-kDa cysteine proteinases. Under reducing conditions, labeling was associated with four similar polypeptides (29–34 kDa), which were also detected when incubation with Z-Tyr[¹²⁵I]-AlaCHN₂ was carried out after cell lysis. Labeling was completely abolished if lysates were first incubated with 20 M E-64 and then exposed to the¹²⁵I-tagged inhibitor, thus confirming the specificity of the compound toward cysteine proteinases.     

Morphological peculiarities of juxtaglomerular apparatus of the kidney in rats with hereditary stress-induced arterial hypertension (NISAG rats)

Structure of juxtaglomerular apparatus of the kidney in NISAG rats and morphometric parameters of renin-producing juxtaglomerular, cells of afferent arterioles attest to its activation. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 127, No. 5, pp. 576–579, May, 1999     

Vitellocyte ultrastructure in the cestode Didymobothrium rudolphii (Monticelli, 1890): possible evidence for the recognition of divergent taxa within the Spathebothriidea

In the spathebothriidean tapeworm Didymobothrium rudolphii (Monticelli, 1890) the fine structure of the vitellocytes at different stages of their development within the vitelline follicles, vitelline ducts and uterus was studied for the first time using transmission electron microscopy. The vitellocyte inclusions of D. rudolphii are shell globule clusters containing tightly packed shell globules associated with a matrix of moderate electron density, glycogen granules, large electron-lucent lipid droplets (up to 3 μm in diameter), and, occasionally, a lipid droplet may occur in the nucleus of the vitellocytes. The diameter of the clusters ranges from 0.4 to 2.5 μm, the number of shell globules in the clusters varies from 8 to 45, and the size of the globules ranges from 0.12 to 0.25 μm and they are of approximately homogeneous sizes within a cluster. Most vitellocyte lipid droplets have a heterogeneous configuration with a ‘cavity’ inside them when they are within vitelline ducts and intrauterine eggs. Vitellocytes of the eggs contain dark concentric bodies and lipid droplets. The interstitial tissue has a syncytial structure. The morphological parameters of the diameter and shape of shell globule clusters, arrangement of shell globules in clusters, number and diameter of globules within clusters, types of lipid droplets and presence of dark concentric bodies are compared with those of two other spathebothriidean genera, Cyathocephalus and Diplocotyle. The comparative data demonstrate that vitelline material morphology has unique features in three spathenothriidean genera and may be used as evidence for the recognition of separate taxa.     

Distribution of acetylcholinesterase activity during the development of cysticercoids of Hymenolepis diminuta (Cestoda)

The distribution of acetylcholinesterase (AChE) in oncospheres and developing cysticercoids of Hymenolepis diminuta was examined. The enzyme was localized in the nervous system and in some non-nerve cells of these larvae. In oncospheres AChE was detected in hook muscles and in the binucleated medullar center that is known to enclose two neurons. At early developmental stages of the cysticercoids the enzyme was localized in the post-oncospheral hook muscles and in subtegumental muscle fibers of the cercomer. At medium and late stages of development the activity of AChE was detected in the developing nervous system and in two and, subsequently, in four populations of cells, which gradually spread over the whole internal wall of the cyst, thus forming a thin multilayer AChE-positive lining of the cyst cavity. Following withdrawal of the scolex the lining separates the parenchyma of the turned neck from the cyst tissues and remains AChE-positive during the whole life of the parasite, i.e. up to the death of the infected host. The role played by non-neural AChE associated with the cyst cavity lining is unknown, but seems to regulate both the transport of nutrients and minerals into the scolex and waste substances in the opposite direction.     

Spermiogenesis and sperm ultrastructure of the pseudophyllidean cestode Triaenophorus nodulosus (Pallas, 1781)

Spermiogenesis and ultrastructure of the spermatozoon of the pseudophyllidean cestode Triaenophorus nodulosus (Pallas, 1781), a parasite of pike Esox lucius, has been studied by transmission electron microscopy. Spermiogenesis involves firstly the formation of a zone of differentiation with two centrioles associated with striated roots, and an intercentriolar body between them, subsequent growth of the two flagella of unequal length, and a formation of a median cytoplasmic process exhibiting patches of dense material. The nucleus penetrates into spermatid body after flagellar rotation and proximo-distal fusion has started. The mature spermatozoon of T. nodulosus is filiform and contains two axonemes of 9+“1” pattern of the Trepaxonemata, nucleus, cortical microtubules parallel to the spermatozoon axis, and electron-dense granules. The anterior extremity of the gamete contains a single centriole surrounded by numerous electron-dense tubular structures exhibiting spiral arrangement and giving rise to lateral projections, which correspond to the crested body. When the crested body disappears, the spiral pattern of electron-dense tubular structures is changed into a ring, persisting until the centriole of the second axonemes appears. This structure of the crested body of T. nodulosus is unique among the Eucestoda.     

A new model of the depressive syndrome induced by administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in rats

Animals treated with MPTP neurotoxin displayed lowered motor and exploratory activity in the open field test, reduced daily intake of water with a preference for sugar solution over water, prolonged immobilization, and increased index of depression in the forced swimming test. The changes in rat behavior were preserved for at least a week after withdrawal of the drug. The data attest to the development of a state of lowered motivational activity combined with ahedony and “behavioral despair” in response to MPTP, making it possible to consider this state as a new experimental model of dopamine-dependent depressive syndrome in rats. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N ^o 2, pp. 125–128, February, 1995     

Novel therapies of multidrug-resistant Pseudomonas aeruginosa and Acinetobacter spp . infections: the state of the art

Gram-negative non-fermenting bacilli, particularly Pseudomonas aeruginosa and Acinetobacter spp., are important opportunistic pathogens in hosppitalized patients, contributing to their morbidity and mortality. Recently, a rapid increase in frequency of multidrug-resistant clinical strains is being recorded, making the available therapeutic options very limited. Apart from the development of novel classes of antimicrobials, there is renewed interest in the use of old agents or new combinations of available drugs. Numerous in vitro investigations have been reported on the efficacy of different antimicrobials; however, they should be evaluated in experimental infection models and clinical trials. Novel approaches are being investigated, such as inhibition of virulence factor expression by pathogens or inhibition of their metabolic pathways. The use of bacteriophages, particularly those genetically modified, remains an alternative option in the therapy of infections caused by multidrug-resistant strains. Several vaccines against P. aeruginosa are under development. Apart from therapy with antimicrobial agents, eradication of outbreaks comprises implementation of strict infection control measures and prudent use of antimicrobials.     

Records of cysticercoids of Fimbriarioides tadornae Maksimova, 1976 and Branchiopodataenia gvozdevi (Maksimova, 1988) (Cyclophyllidea, Hymenolepididae) from brine shrimps at the Mediterranean coasts of Spain and France, with a key to cestodes from Artemia spp. from the Western Mediterranean

Cysticercoids of two cestode species isolated from brine shrimps from the Mediterranean coasts of Spain and France are described. Fimbriarioides tadornae Maksimova, 1976 (adults known as parasites of Tadorna tadorna) was recorded from Artemia parthenogenetica and A. franciscana from Spain (Bras del Port and River Ebro Delta, respectively), and from A. franciscana from Aigues-Mortes, France (new geographical record). Branchiopodataenia gvozdevi (Maksimova, 1988) (adults known as parasites of Larus genei) was recorded from A. parthenogenetica (Bras del Port, Spain), A. salina (San Pedro del Pinatar, Spain) and A. franciscana (River Ebro Delta, Spain); this is the first record of the species in Europe. An illustrated key to cysticercoids of 12 cestode species from Artemia spp. from the Western Mediterranean is proposed.     

Cardioprotective activity of alcoholic extract of Tinospora cordifolia (Willd.) Miers in calcium chloride-induced cardiac arrhythmia in rats

The present study investigated the antiarrhythmic activity of alcoholic extract of Tinospora cordifolia (T.cordifolia) in CaCl 2 induced arrhythmia.CaCl 2 (25 mg/kg) was administered by intravenous infusion (iv) to produce arrhythmia in rats.The animals were then treated with T.cordifolia extract (150,250,and 450 mg/kg) and verapamil (5 mg/kg,iv).Lead II electrocardiogram was monitored.Plasma calcium,sodium and potassium levels were measured.In CaCl 2 induced arrhythmia,heart rate was decreased by 41.10%,T.cordifolia at 150,300,and 450 mg/kg decreased the heart rate by 26.30%,29.16%,and 38.29%,respectively,and verapamil reduced the heart rate by 9.70% compared to the normal group.The PQRST waves were normalized and atrial and ventricular fibrillation was controlled in rats treated with verapamil and T.cordifolia.CaCl 2 increased calcium and sodium levels and decreased potassium levels in blood.T.cordifolia dose-dependently decreased calcium and sodium levels and increased potassium levels.Hence,T.cordifolia can be used in antiarrhythmic clinical settings and beneficial in atrial and ventricular fibrillation and flutter and may be indicated in ventricular tachyarrhythmia.     

Enkephalinase mechanisms of resistance and tolerance to the analgesic action of morphine in rats. II. Differential effects of naloxone in morphine-sensitive,morphine-resistant,and morphine-tolerant rats

Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 116, N ^o 7, pp. 6–9, July, 1993.     

Laying the Cytotaxonomic Foundations of a New Model Grass, Brachypodium distachyon (L.) Beauv.

Brachypodium distachyon is a ubiquitous, temperate grass species which is being developed and exploited as an alternative model to rice, in order to gain access to important syntenic regions of the genomes of less tractable relatives such as wheat. As part of this initiative, this paper describes for the first time the cytotaxonomy of members of the polyploid series of this species, and challenges the assumption that the series evolved simply by chromosome doubling. In situ hybridization using genomic DNA probes and rDNA markers uncovers a hybrid origin of several of the polyploid ecotypes, and sheds light upon the complex evolution of this species and its close relatives.