Evidence of DNA rearrangements in the 128-kilobase pBtoxis plasmid of Bacillus thuringiensis israelensis

BUPM97 is a novel Tunisian isolate of Bacillus thuringiensis israelensis presenting insecticidal activity against Culex pipiens larvae. The δ-endotoxins pattern of this strain was different from that of the reference strain B. thuringiensis israelensis H14. Therefore, the study of its cry genes content was carried out by restriction-fragment-length-polymorphism (RFLP) using specific cry genes probes and by DNA sequencing. It was clearly demonstrated that in the strain BUPM97 the cry4A and cry10A genes were deleted from the B. thuringiensis israelensis 128-kb pBtoxis plasmid. In addition, a strong DNA sequence polymorphism was evidenced in the same plasmid downstream from the cry4B gene. This very particular DNA dynamic evidenced in this new strain of B. thuringiensis israelensis should be taken into consideration, regarding the strain stability during the industrial production of B. thuringiensis bioinsecticides.     

An ultrastructural study of oogenesis in the planarian Schmidtea mediterranea (Platyhelminthe, Paludicola)

The ovary of the freshwater planarian Schmidtea mediterranea has been studied for the first time using both light and electron microscopy methods. The ultrastructure of the ovary revealed two types of cells: accessory cells and germinal cells at various stages of differentiation, distributed along a maturation axis. Initially, oogonia underwent cytoplasm growth due to the development of organelles, such as endoplasmic reticulum, Golgi complex, and mitochondria, which are all involved in the production of cytoplasmic inclusions or yolk globules. It is shown that the chromatoid body and fibrogranular aggregates may participate in the synthesis of vitelline inclusions. When completely mature, the oocytes have become larger, due to the accumulation of nutritive inclusions, which are round in shape and have a paracrystalline structure. These inclusions are interpreted as being yolk globules and may represent a kind of nutritive material for the developing embryo. These ultrastructural features of the ovary agree with the available phylogenetic tree, based on morphological and karyological characters that considers Schmidtea group as a genus and not a subgenus. The presence of sperm between the oocytes suggests that fertilization may occur within the ovary, representing an uncommon condition within the Triclads, in which fertilization usually takes places outside of the ovaries.     

Comparison of different delivery systems of DNA vaccination for the induction of protection against tuberculosis in mice and guinea pigs

The great challenges for researchers working in the field of vaccinology are optimizing DNA vaccines for use in humans or large animals and creating effective single-dose vaccines using appropriated controlled delivery systems. Plasmid DNA encoding the heat-shock protein 65 (hsp65) (DNAhsp65) has been shown to induce protective and therapeutic immune responses in a murine model of tuberculosis (TB). Despite the success of naked DNAhsp65-based vaccine to protect mice against TB, it requires multiple doses of high amounts of DNA for effective immunization. In order to optimize this DNA vaccine and simplify the vaccination schedule, we coencapsulated DNAhsp65 and the adjuvant trehalose dimycolate (TDM) into biodegradable poly (DL-lactide-co-glycolide) (PLGA) microspheres for a single dose administration. Moreover, a single-shot prime-boost vaccine formulation based on a mixture of two different PLGA microspheres, presenting faster and slower release of, respectively, DNAhsp65 and the recombinant hsp65 protein was also developed. These formulations were tested in mice as well as in guinea pigs by comparison with the efficacy and toxicity induced by the naked DNA preparation or BCG. The single-shot prime-boost formulation clearly presented good efficacy and diminished lung pathology in both mice and guinea pigs.     

Oogenesis of Perinereis macropus Claparede, 1870 (Annelida, Polychaeta) in the Gulf of Gabes (Tunisia)

The polychaete Perinereis macropus (Claparède, 1870) (Nereididae) is present in the Gulf of Gabes, but its reproductive biology is unknown. An intensive study was conducted from August 2004 to July 2005 to characterize the life cycle of a population in the mouth of wadi Ferd in Gabes. The examination of sexual products allows us to describe the morphological oocytes aspects at different stages. The results show that P. macropus has an asynchronous oogenesis. Moreover, the biometric study of oocytes growth allows us to clarify the female sexual cycle. The female maturity occurs in April and the mature oocyte diameter is approximately 250+/-32.67 microm. However, spawning occurs in May, when the seawater temperature starts rising. We propose that the reproductive season stretches from March to June in the Gulf of Gabes.     

Morphological characteristics of the pharynx of Nereis diversicolor Müller, 1776 (Polychaetes, Nereidae)

Within the biometrical study of the Nereidae and in order to characterize the different populations, we looked for the most relevant biometrical characteristics. In fact, for these sea-animals, the weight and length remain just significant statements within certain limits; on the other hand the length of the maxillary, the number of paragnaths which decorate the proboscis can be precisely known.     

The effects of Ramadan intermittent fasting on the underlying mechanisms of force production capacity during maximal isometric voluntary contraction

The aim of the present study was to investigate the effects of Ramadan intermittent fasting (RIF) on the underlying mechanisms of force production capacity during maximal voluntary isometric contraction (MVIC) using the superimposed twitch technique. Ten healthy male physical education students performed three MVIC of the knee extensor superimposed with nerve electrical stimulation during four testing phases: one week before Ramadan (BR), at the end of the first week of Ramadan (R-1), during the fourth week of Ramadan (R-4) and two weeks after Ramadan (AR). This study was performed during Ramadan 2016. MVIC values, voluntary activation level (VAL), potentiated resting twitch and electromyography signals were recorded during each MVIC. The French version of the Profile of Mood States questionnaire (POMS-f) was used to evaluate the subjective mood states in each testing session. The results showed that MVIC values (890.57 ± 67.90 vs. 816.46 ± 54.72 N) and VAL (87.73 ± 3.27 vs. 77.32 ± 7.87%) decreased at R-1 compared to BR (p < 0.001). However, the neuromuscular efficiency and the potentiated resting twitch remained unchanged during Ramadan (R). Results showed that depression (p < 0.01; 6.3 ± 1.57 vs. 4.7 ± 1.25), fatigue (p < 0.001; 9.2 ± 1.93 vs. 4.6 ± 2.01) and anxiety (p < 0.001; 6.4 ± 1.51 vs. 11.8 ± 1.23) scores of POMS-f were higher during R-1 compared to BR. In conclusion, RIF-related impairment of maximal muscle force seems to be related to nervous alterations of the VAL, whereas the RIF did not adversely affect peripheral mechanisms.

Abbreviations’ List: ANOVA: Analysis of variance; AR: After Ramadan; BMI: Body-mass index; BR: Before Ramadan; EMG: Electromyography; ER: End of Ramadan; MF: Mean frequency; M_max: Peak-to-peak M-wave amplitudes; MVIV: Maximal voluntary isometric contraction; NES: Nerve electrical stimulation; NME: Neuromuscular efficiency; POMS-f: French version of the Profile of Mood States questionnaire; R: Ramadan; R-1: First week of Ramadan; R-4: Fourth week of Ramadan; RF: Rectus femoris; RIF: Ramadan intermittent fasting; RMS: Root mean square; VAL: Voluntary activation level; VL: Vastus lateralis; VM: Vastus medialis.     

High prevalence of the point mutation in exon 6 of the xeroderma pigmentosum group A-complementing (XPAC) gene in xeroderma pigmentosum group A patients in Tunisia.

Xeroderma pigmentosum (XP) patients in Tunisia who belong to the genetic complementation group A (XPA) have milder skin symptoms than do Japanese XPA patients. Such difference in the clinical features might be caused by the difference in the site of mutation in the XP A-complementing (XPAC) gene. The purpose of this study is to identify the genetic alterations in the XPAC gene in the Tunisian XPA patients and to investigate the relationship between the clinical symptoms and the genetic alterations. Three sites of mutation in the XPAC gene have been identified in the Japanese XPA patients, and about 85% of them have a G-->C point mutation at the splicing acceptor site of intron 3. We found that six (86%) of seven Tunisian XPA patients had a nonsense mutation in codon 228 in exon 6, because of a CGA-->TGA point mutation, which can be detected by the HphI RFLP. This type of mutation is the same as those found in two Japanese XPA patients with mild clinical symptoms. Milder skin symptoms in the XPA patients in Tunisia than in those in Japan, despite mostly sunny weather and the unsatisfactory sun protection in Tunisia, should be due to the difference in the mutation site.     

Evidence of the Importance of the Met115 for Bacillus thuringiensis subsp. israelensis Cyt1Aa Protein Cytolytic Activity in Escherichia coli

Cyt1Aa is a cytolytic toxin, found together with the delta-endotoxins in Bacillus thuringiensis subsp. israelensis parasporal insecticidal crystals. The latter are used as an environmental friendly insecticide against mosquitoes and black flies. Contrary to Cry delta-endotoxin, the mode of action of Cyt1Aa is not completely understood. In the absence of direct structural data, a novel mutated cyt1Aa gene was used to obtain indirect informations on Cyt1Aa conformation changes in the lipid membrane environment. A mutated cyt1Aa gene named cyt1A97 has been isolated from a B. thuringiensis israelensis strain named BUPM97. The nucleotide sequence predicted a protein of 249 amino acids residues with a calculated molecular mass of 27 kDa. Both nucleotide and amino acid sequences similarity analysis revealed that cyt1A97 presents one amino acid different from the native cyt1Aa gene. This mutation was located in the helix α C corresponding to a substitution of Met¹¹⁵ by a Thr. The heterologous expression of the cyt1A97 and another cyt1Aa-type gene called cyt1A98, not affected by such mutation used as control, was performed in Escherichia coli. It revealed that the mutated Cyt1A97 protein was over produced as inclusion bodies showing a very weak toxicity to E. coli contrarily to Cyt1A98 that stopped E. coli growth. Hence, hydrophobic residue Met at position 115 of Cyt1Aa should play a very important role for the maintenance of the structure and cytolytic functions of Cyt1Aa.     

The freshwater planarian Dugesia sicula Lepori, 1948 (Platyhelminthes, Tricladida) in Tunisia: ecology, karyology, and morphology

A study was carried out on freshwater planarians of the genus Dugesia from three localities in northern Tunisia: Joumine, Chiba, and Lebna. The three populations are fissiparous under field conditions and do not possess reproductive organs, except for some cases of hyperplastic ovaries in the Joumine population. After five months of breeding under laboratory conditions, with water temparature between 18 and 25?°C and under short photoperiod, 8.33 to 26% of specimens from the Joumine stream became sexualized, together with considerable increase of body length and width. For the other two populations the sexualization rate was very small. Extended period of darkness apparently played a positive role in this sexualization. The phenomenon of sexualization is discussed in the context of reproductive strategies. Histological and karyological studies of the sexualized animals allowed identification of these usually fissiparous planarians as representatives of Dugesia sicula Lepori, 1948. The species is here reported for the first time from Tunisia. The populations are restricted to permanent water basins resulting from the construction of dams in large rivers.