Molecular Detection of Chlamydia trachomatis and Other Sexually Transmitted Bacteria in Semen of Male Partners of Infertile Couples in Tunisia: The Effect on Semen Parameters and Spermatozoa Apoptosis Markers

This study was undertaken to determine the prevalence of Chlamydia trachomatis, Mycoplasmas, and Ureaplasmas in semen samples of the male partners of infertile couples and to investigate whether Chlamydia trachomatis could initiate apoptosis in human spermatozoa. A total of 85 males partners of infertile couples undergoing routine semen analysis according to World Health Organization guidelines were included. Specimens were examined for the presence of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma urealyticum and Ureaplasma parvum by Real time PCR (qPCR). Semen specimens were analysed for the appearance of apoptotic markers (sperm DNA fragmentation, activated caspase 3 levels, mitochondrial membrane potential (ΔΨm)) using flow cytometry. C. trachomatis, N. gonorrhoeae, U. urealyticum, M genitalium were detected in semen samples of 13 (15.2%), 5 (5.8%), 5 (5.8%) and 3 (3.5%) male partners of infertile couples, respectively. M. hominis and U. parvum were detected in semen sample of only one patient (1.1%). The semen of infertile men positive for C. trachomatis showed lower mean of semen count and lower rapid progressive motility (category [a]) of spermatozoa compared to uninfected men with statistically significances (p = 0.02 and p = 0.04, respectively). Flow cytometry analyses demonstrated a significant increase of the mean rate of semen with low ΔΨm and caspase 3 activation of infertile men positive for C. trachomatis compared to uninfected men (p = 0.006 and p = 0.001, respectively). DNA fragmentation was also increased in sperm of infertile men positive for C. trachomatis compared to uninfected men but without statistical significances (p = 0.62). Chlamydial infection was associated to loss of ΔΨm and caspase 3activation. Thus, C. trachomatis infection could be incriminated in apoptosis induction of spermatozoa. These effects may explain the negative direct impact of C. trachomatis infection on sperm fertilizing ability.     

Notes on the Husbandry and Long‐Term Transportation of Bull Ray (Pteromylaeus bovinus) and Dolphinfish (Coryphaena hippurus and Coryphaena equiselis)

Bull rays (Pteromylaeus bovinus) and Dolphinfish (Coryphaena hippurus and Coryphaena equiselis) were collected in Olhão (south of Portugal). These animals hosted multiple parasites, namely Caligus spp., and underwent a variety of treatments to remove them. Of all treatments tested, hydrogen peroxide showed the best results, although only concentrations above 100 ppm were effective in parasite removal. These high concentrations, however, proved to be highly toxic for the fish and led to the loss of some animals, especially those which had been handled before treatment. A total of 14 Bull rays were transported to Bolougne‐Sur‐Mer (France) by road and some animals were lost, which was attributed to excessive time in transit (>45 hr). In another transport, three Bull rays and 10 Dolphinfishes were moved to Stralsund (Germany) by road and air. The mechanical wounds suffered by one of the Bull rays during transport led to its death and, consequently, a deterioration of water quality in the tank containing two other conspecifics. This deterioration of water quality resulted in problems for the other two Bull rays, and one perished approximately 48 hr after arrival. The authors concluded that Dolphinfish can be transported with a low bioload for at least 27 hr, and Bull rays should not undergo transports longer than 35 hr. Special attention must be taken to injured animals, since this can lead to a decrease in water quality and consequently affect other animals in the same transport tank. Zoo Biol. 32:222–229, 2013. © 2012 Wiley Periodicals, Inc.     

Antimicrobial Activity of Essential Oil Components Against Potential Food Spoilage Microorganisms

The antimicrobial activity of six essential oil components against the potential food spoilage bacteria Aeromonas (A.) hydrophila, Escherichia (E.) coli, Brochothrix (B.) thermosphacta, and Pseudomonas (P.) fragi at single use and in combination with each other was investigated. At single use, the most effective oil components were thymol (bacteriostatic effect starting from 40 ppm, bactericidal effect with 100 ppm) and carvacrol (50 ppm/100 ppm), followed by linalool (180 ppm/720 ppm), α-pinene (400 ppm/no bactericidal effect), 1,8-cineol (1,400 ppm/2,800 ppm), and α-terpineol (600 ppm/no bactericidal effect). Antimicrobial effects occurred only at high, sensorial not acceptable concentrations. The most susceptible bacterium was A. hydrophila, followed by B. thermosphacta and E. coli. Most of the essential oil component combinations tested showed a higher antimicrobial effect than tested at single use. Antagonistic antimicrobial effects were observed particularly against B. thermosphacta, rarely against A. hydrophila. The results show that the concentration of at least one of the components necessary for an antibacterial effect is higher than sensorial acceptable. So the use of herbs with a high content of thymol, carvacrol, linalool, 1,8-cineol, α-pinene or α-terpineol alone or in combination must be weighted against sensorial quality.     

<Emphasis Type="Italic">Pseudodiplectanum syrticum</Emphasis> n. sp. (Monogenea: Diplectanidae), a parasite of <Emphasis Type="Italic">Synapturichthys kleinii</Emphasis> (Teleostei: Soleidae) from off Tunisia

Pseudodiplectanum syrticum n. sp. (Monogenea: Diplectanidae) is described from the gills of Synapturichthys kleinii (Risso) collected from the Gulf of Gabès in the Mediterranean Sea off Tunisia. The new species differs from the congeneric species by the morphology of the penis and vagina. It resembles the Mediterranean species P. kearni Vala, Lopez-Roman & Boudaoud, 1980 from Solea vulgaris Quensel and P. gibsoni Oliver, 1987 from Michrochirus variegatus (Donovan) in its tubular penis, but it differs from these two species by having a vagina with a long tube presenting two loops and a male copulatory organ ending in a curved point. It also differs in the morphology and size of the transverse bars of the haptor. In P. syrticum the ventral transverse bar and dorso-lateral bars are longer than those of P. gibsoni (114 vs 69.5-89 microm and 48 vs 33-44 microm, respectively).     

Morphological and functional evaluation of angiogenesis of a xenografted human sarcoma in nude mice

The implantation of tumour cells in normal tissues and the subsequent induction of angiogenesis by the growing xenograft were studied by means of immunohistochemistry and digital image analysis. Tumour growth was induced by injection of a human spindle cell sarcoma (ES3) into the subcutis of HsdCpb:NMRI-nu/nu mice. In vivo injection of Hoechst 33342 was used as a marker of perfusion. The vasculature was stained with specific antibodies and subsequently analysed by digital image analysis. Starting at day 3 up to day 6, angiogenesis could be detected and the relative amount of perfusion within the investigated area reached a peak at day 6. This method, which allows investigation of both functional and morphometric characteristics of human xenograft vasculature, serves as an excellent assay for evaluation of antiangiogenic therapies in translational research of experimental tumours.     

Identification of Important Amino Acid Residues of the Na+-Ca2+ Exchanger Inhibitory Peptide,XIP

The Na⁺-Ca²⁺ exchanger plays an important role in cardiac contractility by moving Ca²⁺ across the plasma membrane during excitation-contraction coupling. A 20 amino acid peptide, XIP, synthesized to mimic a region of the exchanger, inhibits exchange activity. We identify here amino acid residues important for inhibitory function. Effects of modified peptides on Na⁺-Ca²⁺ exchange activity were determined. Exchange activity was assessed as ⁴⁵Ca²⁺ uptake into Na⁺-loaded cardiac sarcolemmal vesicles. We find that the entire length of XIP is important for maximal potency, though the major inhibitory components are between residues 5 and 16. Basic and aromatic residues are most important for the inhibitory function of XIP. Substitutions of arginine 12 and arginine 14 with alanine or glutamine dramatically decrease the potency of XIP, suggesting that these residues play a key role in possible charge-charge interactions. Substitutions of other basic residues with alanines or glutamines had less effect on the potency of XIP. All aromatic residues participate in binding with the exchanger, probably via hydrophobic interactions as indicated by tryptophan fluorescence. A tyrosine is required at position 6 for maximal inhibition and phenylalanine 5 and tyrosine 8 can only be replaced by other aromatic residues. Tyrosine 10 and tyrosine 13 can be replaced with other bulky residues. A specific conformation of XIP, with structural constrains provided by all parts of the molecule, is required for optimal inhibitory function. Received: 19 September 1996/Revised: 20 November 1996     

Über Entwicklungsgeschichte und Ernährungsphysiologie eines neuen niederen Phycomyceten mit Generationswechsel

Ohne ZusammenfassungMit 17 Textabbildungen (21 Einzelbildern).D. 7.     

Analysis of bacterial DNA in synovial tissue of Tunisian patients with reactive and undifferentiated arthritis by broad-range PCR, cloning and sequencing

Introduction

Bacteria and/or their antigens have been implicated in the pathogenesis of reactive arthritis (ReA). Several studies have reported the presence of bacterial antigens and nucleic acids of bacteria other than those specified by diagnostic criteria for ReA in joint specimens from patients with ReA and various arthritides. The present study was conducted to detect any bacterial DNA and identify bacterial species that are present in the synovial tissue of Tunisian patients with reactive arthritis and undifferentiated arthritis (UA) using PCR, cloning and sequencing.

Methods

We examined synovial tissue samples from 28 patients: six patients with ReA and nine with UA, and a control group consisting of seven patients with rheumatoid arthritis and six with osteoarthritis (OA). Using broad-range bacterial PCR producing a 1,400-base-pair fragment from the 16S rRNA gene, at least 24 clones were sequenced for each synovial tissue sample. To identify the corresponding bacteria, DNA sequences were compared with sequences from the EMBL (European Molecular Biology Laboratory) database.

Results

Bacterial DNA was detected in 75% of the 28 synovial tissue samples. DNA from 68 various bacterial species were found in ReA and UA samples, whereas DNA from 12 bacteria were detected in control group samples. Most of the bacterial DNAs detected were from skin or intestinal bacteria. DNA from bacteria known to trigger ReA, such as Shigella flexneri and Shigella sonnei, were detected in ReA and UA samples of synovial tissue and not in control samples. DNA from various bacterial species detected in this study have not previously been found in synovial samples.

Conclusion

This study is the first to use broad-range PCR targeting the full 16S rRNA gene for detection of bacterial DNA in synovial tissue. We detected DNA from a wide spectrum of bacterial species, including those known to be involved in ReA and others not previously associated with ReA or related arthritis. The pathogenic significance of some of these intrasynovial bacterial DNAs remains unclear.