Key role of hydrogen peroxide in antimicrobial activity of spring, Honeydew maquis and chestnut grove Corsican honeys on Pseudomonas aeruginosa DNA.

In honeys, several molecules have been known for their antibacterial or wound healing properties. Corsican honeys just began to be tested for their antimicrobial activity with promising results on Pseudomonas aeruginosa. So, identification of active molecules and their mode of action was determined. Hydrogen peroxide concentrations were evaluated and, in parallel, the minimal inhibitory concentrations (MIC) values were performed with and without catalase. More, the quantity of phenolic compounds and ORAC assay were measured. Observation of antibacterial action was done using scanning electron microscopy (SEM) followed by plasmidic DNA extraction. MIC values of chestnut grove and honeydew maquis honeys vary between 7 and 8%, showing a strong antimicrobial capacity, associated with a plasmidic DNA degradation. When catalase is added, MIC values significatively increase (25%) without damaging DNA, proving the importance of H2 O2 . This hypothesis is confirmed by SEM micrographies which did not show any morphological damages but a depletion in bacterial population. Although, such low concentrations of H2 O2 (between 23 µmol l-1 and 54 µmol l-1 ) cannot explain antimicrobial activity and might be correlated with phenolic compounds concentration. Thus, Corsican honeys seem to induce DNA damage when H2 O2 and phenolic compounds act in synergy by a putative pro-oxidant effect. SIGNIFICANCE AND IMPACT OF THE STUDY: We started to determine the antibacterial efficiency of Corsican chestnut grove and honeydew maquis honeys on Pseudomonas aeruginosa. No morphological alteration of the bacterial surface was observed. Antimicrobial action seems to be related to the synergy between hydrogen peroxide and phenolic compounds. The exerted pro-oxidant activity leads to a degradation of P. aeruginosa plasmidic DNA. This is the first study that investigate the primary antibacterial mechanism of Corsican honeys.

Fasciolosis in the Mediterranean island of Corsica (France): Insights from epidemiological and malacological investigations.

Fasciolosis is a re-emergent parasitic disease of worldwide significance with a major global impact on livestock health and production. In the French Mediterranean island of Corsica, fasciolosis has been recognized for a long time but little is known about its dynamic as the main investigations are outdated. Three compartments - definitive domestic hosts, intermediate hosts and environment - involved in fasciolosis transmission were studied by applying an integrative and extensive approach: (1) farm and abattoir surveys, (2) snail sampling, identification and infection prospection, and (3) snail habitat analysis; and (4) a questionnaire-based survey to inquire about husbandry practices and environmental risks. Our results indicate a significant circulation of the liver flukes in Corsican livestock, with 90% (252/279) of the sampled farms testing positive for anti-F. hepatica antibodies. At the abattoir, 46% (67/149) of cattle were positive for F. hepatica antibodies and eggs were present in the bile of 19% (26/139) bovines. In addition, high prevalence of Dicrocoelium dendriticum (69%) was observed in slaughtered cattle. Malacological surveys registered the occurrence of several lymnaeid species in a variety of habitats throughout the island. In particular, we report for the first time the presence of the invasive lymnaeid snail Pseudosuccinea columella in Corsica, a potential intermediate host for F. hepatica. We also found that the presence of Galba truncatula and, to a lesser extent, that of Peregriana peregra, is associated with altitude. Fasciola hepatica DNA was detected in the latter species occurring at two different sites. Finally, a questionnaire-based study revealed risky management practices among Corsican farmers, low perception of transmission and a suboptimal use of flukicide treatments as main control strategy. Our results show that animal fasciolosis in Corsica is characterised by a significant circulation and a favourable epidemiological scenario for transmission to occur.

Ultrastructural study of spermiogenesis and the spermatozoon of Acanthocephaloides incrassatus (Molin, 1858) (Acanthocephala, Paleacanthocephala, Arhythmacanthidae) from Anguilla anguilla (Pisces, Teleostei) in Urbino ponds (Corsica Island).

This study deals with first ultrastructure features of Acanthocephaloides incrassatus (Paleacanthocephala, Arhythmacanthidae), a parasite of the fish Anguilla anguilla, reported for the first time in a Mediterranean pond. The spermiogenesis of A. incrassatus shows original specificities which have never been pointed out to this day in ultrastructural studies on spermiogenesis: the centriolar derivative is divided into two parts of different densities: an electron-dense, and the other, electron-lucent; a ring form has been observed on each side of the axoneme; a centriole with one central element. After the elaboration of a flagellum of 9+2 pattern, the centriole migrates in a nuclear groove. Rapidly, the centriole disappears. Then, the flagellum migration occurs by a series of processes and gives rise to a spermatozoon. The spermatozoon of A. incrassatus presents its own specificities: it exhibits an evolution of the centriolar derivative characterized by only nine peripheral elements deprived of a central element in the anterior part, then nine peripheral and one central element, and finally, nine peripheral elements with two central elements. An assumption is emitted on a probable correspondence of the evolution of the derivative centriolar during the spermiogenesis and the evolution that occurs in the spermatozoon. Protein granules also show different sizes and forms, full or emptied of their contents compared with data on other Acanthocephala.

Cellular effects induced by Inula graveolens and Santolina corsica essential oils on Staphylococcus aureus.

This study was conducted to evaluate the antibacterial activity of Inula graveolens and Santolina corsica essential oils on Staphylococcus aureus and investigate their effects at the cellular level. The mode of inhibition of both essential oils against S. aureus ATCC 6538P (CIP 53.156) was assessed by determining the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). The effects of time and treatment dose on cell viability were determined by time-kill and bacteriolysis assays. Marked structural changes were observed by transmission electron microscopy (TEM). A bactericidal mode of inhibition was established for both essential oils, which rapidly reduced the cell viability of S. aureus at their MIC (5 mg.ml(-1)). No lysis occurred after treatments with the MIC and eight times the MIC of each essential oil. Invaginations of the plasmic membrane with thickenings of the cell wall as well as an aggregation of the cytoplasmic contents were observed in S. aureus cells treated with the MIC of both essential oils. These results suggest that the cytoplasmic membrane and the cell wall are involved in the toxic action of Inula graveolens and Santolina corsica essential oils.

Sperm ultrastructure of the digenean Siphoderina elongata (Platyhelminthes, Cryptogonimidae) intestinal parasite of Nemipterus furcosus (Pisces, Teleostei).

The ultrastructure of the spermatozoon of Siphoderina elongata was studied by transmission electron microscopy. A description and drawings of the mature spermatozoon are presented in this paper. Several ultrastructural elements of this male gamete have been observed: a nucleus, two mitochondria, two axonemes of 9 + "1" pattern, external ornamentation of the plasma membrane, spine-like bodies and cortical microtubules. The presence, the location or the number of these elements have been compared with other digenean spermatozoa. Moreover, a close attention was paid to the organization of the external ornamentation region. This zone presents a single row of cortical microtubules disposed in a semi-circle around a mitochondrion and associated with external ornamentation and spine-like bodies. The aim of this study is to highlight criteria which can be interesting in Platyhelminthes phylogeny.

Comparison of arsenic and antimony contents in tissues and organs of brown trout caught from the river Presa polluted by ancient mining practices and from the river Bravona in Corsica (France): a survey study.

The present study evaluates the concentrations of arsenic (As) and antimony (Sb) in the intestine, liver, muscle, gonads, gills, and kidney of Salmo trutta subsp. from the Presa River in Corsica (France; n = 10), which crosses an abandoned arsenic mine, and from the Bravona River (reference site; n = 10). Both metalloids were analyzed by means of ICP-MS. The relationships between fish size (length and weight) and metalloid concentrations in their tissues were investigated by linear regression analysis. In all fish samples concentrations of As and Sb (expressed as micrograms per gram fresh weight) were highest in the kidney. Lowest Sb concentrations were found in the muscle, whereas lowest As concentrations were found in the gonads of S. trutta. Two organotropisms were revealed: one for As-kidney (21.4656) > intestine (3.9535) > gills (3.0404) > liver (1.1743) > muscle (0.9976) > gonads (0.8081); and the other for Sb-kidney (0.70067) > gills (0.6181) > intestine (0.2576) > gonads (0.1673) > liver (0.9625) > muscle (0.0753). Results of linear regression analysis in most cases showed a significant negative correlation between metalloid concentration and fish size. Highly significant (p < 0.05) negative correlations were found between fish length and As concentration in the gonads, as well as between fish length and Sb concentrations in the gills. Arsenic concentrations in female fish were significantly higher than those in males in the kidney, gonads, gills, and liver. The same results were found for Sb, except in the liver, where the tendency was reversed.

Molecular investigation of tick-borne pathogens in ixodid ticks infesting domestic animals (cattle and sheep) and small rodents (black rats) of Corsica, France.

Although livestock farming (sheep, goats, pigs, and cattle) is an important economic activity in Corsica, a French Mediterranean island, knowledge about the tick fauna and microorganisms carried by them remains scarce. This study aimed to investigate the presence and perform molecular characterization of Anaplasmataceae, Rickettsia spp., and Borrelia burgdorferi sensu lato (sl) in tick species collected in Corsica. Ticks from cattle (Bos taurus), sheep (Ovis aries), and rodents (Rattus rattus) were collected from May to September 2016. DNA was purified from ticks, submitted to quantitative real-time polymerase chain reaction (qPCR) and sequenced for phylogenetic analysis. In total, 660 ticks were collected from 111 animals during the study. The most abundant collected tick species from cattle was Rhipicephalus bursa (n = 495; 84.5%), followed by Hyalomma marginatum (n = 91; 15.5%). Rhipicephalus bursa and Ixodes ricinus were the only tick species collected from sheep and rodents, respectively. Overall, Rickettsia was the most common pathogen group (n = 48; 24%) detected in ticks. Sequence analysis of partial gltA and ompA genes revealed the presence of Ri. aeschlimannii and Candidatus Ri. barbariae. Anaplasmataceae DNA was detected in eight (6%) of the 127 cattle pools and in one (2%) of the 61 R. bursa specimens collected from sheep. Sequence analysis of the rpoB gene revealed the presence of one Anaplasma species, A. marginale. Borrelia burgdorferi sl DNA was detected in one pool of H. marginatum collected from cattle and in two (15%) of the 13 I. ricinus pools collected from nine black rats. To our knowledge, this is the first report of the occurrence and molecular characterization of Candidatus Ri. barbariae, an emerging member of the Rickettsia group causing spotted fever, in Corsica. The detection of B. burgdorferi sl DNA, which was previously believed to be rare in Corsica, confirms the presence of this agent on the island.

Parasitofauna study of the brown trout, Salmo trutta (Pisces, Teleostei) from Corsica (Mediterranean island) rivers.

Corsica is a Mediterranean island characterised by a great number of rivers. Salmonides are the main fishes which populate these rivers. Very appreciated by fishermen, Salmonides are represented by three species in the insular hydrographical network, among which an autochthonous species, the brown trout (Salmo trutta). In the present work, we have analysed the parasitofauna of this species. According to our knowledge, this research has never been carried out in Corsica. In a first step, we drew up an inventory of the parasites found in this freshwater fish. In a second step, we studied the differences which appeared in the composition of parasite communities of this species.

Ultrastructure of the spermatozoon of Helicometroides atlanticus (Digenea, Monorchiidae), an intestinal parasite of Parapristipoma octolineatum (Pisces, Teleostei) in Senegal.

The mature Helicometroides atlanticus spermatozoon exhibits an anterior electron-dense material, two axonemes of the 9+"1" pattern, a nucleus, two mitochondria, extramembranous ornamentations located at the level of the first mitochondrion and cortical microtubules arranged into two fields: a ventral field corresponding to the mitochondrial side and a dorsal field corresponding to the nuclear side. It lacks of spine-like body and a cytoplasmic expansion as in Monorchis parvus the only species of Monorchioidea which spermatozoon has been described until now. Nevertheless, it is distinguished from M. parvus by the presence of an anterior electron-dense material and two axonemes which appear one after the other.

Effect of duration of feed withdrawal and transportation time on muscle characteristics and quality in Friesian-Holstein calves.

Twenty-week-old Friesian-Holstein calves were used to assess the influences of the duration of feed withdrawal before transport (1 or 11 h) and of transport time (1 or 11 h) on carcass and muscle characteristics and meat sensory qualities. One hundred twelve calves were used for live weight and carcass measurements, following a 2 x 2 factorial design (28 replicates). Twelve calves were randomly selected in each treatment group to examine muscle characteristics and sensory quality of longissimus lumborum (LL) and semimembranosus (SM). Long transport (11 h) increased loss in live weight (P < .001) and dressing percentage (P < .001). Feed withdrawal for 11 h also increased dressing percentage (P < .001). Long transport resulted in decreased liver weight (P < .05), glycolytic potential (P < .01), and pH at 4 h postmortem (P < .05) in the LL. Drip loss, compositional traits, cooking loss, and sarcomere length in the LL, as well as drip loss, pH values, compositional traits, cooking loss, and sarcomere length in the SM, were unaffected by the treatments. However, long transport decreased tenderness score in the LL (P < .05) and SM (P < .01). It also increased myofibrillar resistance (P < .001) in the latter muscle. The results demonstrate an unfavorable effect of long transport on the sensory quality of veal. This effect cannot be explained on the basis of differences in pH values and(or) compositional characteristics.

Sperm ultrastructure of the digenean Aphallus tubarium (Rudolphi, 1819) Poche, 1926 (Platyhelminthes, Cryptogonimidae) intestinal parasite of Dentex dentex (Pisces, Teleostei).

The ultrastructural organization of the spermatozoon of a cryptogonimid digenean, Aphallus tubarium, a parasite of Dentex dentex, is described. The spermatozoon possesses the elements found in other digeneans: two axonemes with 9+"1" pattern, a mitochondrion, a nucleus, cortical microtubules, external ornamentation and spine-like bodies. However, the mitochondrion appears as a cord with a bulge; this characteristic has never been described in other studied cryptogonimid and in other digeneans except in one lepocreadiid, Holorchis micracanthum. Likewise, the presence of a thin cytoplasm termination in the anterior part of the spermatozoon has never been pointed out in the cryptogonimids.

Ultrastructural study of the male gamete of Glossobothrium sp. (Cestoda: Bothriocephalidea: Triaenophoridae) a parasite of Schedophilus velaini (Perciformes: Centrolophidae) in Senegal.

This paper describes the ultrastructure of the male gamete of Glossobothrium sp. (Bothriocephalidea: Triaenophoridae). The mature spermatozoon of Glossobothrium sp. is filiform and possesses two axonemes, a single helicoidal crested body, a parallel nucleus, parallel cortical microtubules and granules of glycogen. In Glossobothrium sp. we describe for first time a 200-250 nm thick crest-like body in the Bothriocephalidean. The anterior part of the spermatozoon exhibits a ring of 27 electron-dense cortical microtubules encircling the first axoneme. This structure persists until the appearance of the second axoneme. When the ring of electron-dense cortical microtubules disappears, the spermatozoon exhibits two bundles of thin cortical microtubules. The posterior part of the spermatozoon contains the posterior extremity of the second axoneme, the posterior extremity of the nucleus and few cortical microtubules. Soon nucleus disappears and the axoneme is disorganized. Thus the posterior extremity of the spermatozoon of Glossobothrium sp. exhibits only singlets produced by the disorganization of the doublets of the second axoneme and few cortical microtubules. This type of posterior extremity of the mature spermatozoon has never been described previously in the Triaenophoridae.

Ultrastructural study of spermiogenesis and the spermatozoon of Cavisoma magnum (Southwell, 1927) (Acanthocephala, Palaeacanthocephala, Cavisomidae), from Siganus lineatus (Pisces, Teleostei, Siganidae) (Valenciennes, 1835) in New Caledonia.

This paper presents an ultrastructural study of Cavisoma magnum (Acanthocephala, Cavisomatidae) with a Transmission Electron Microscopy tool. This parasite of the fish Siganus lineatus is here reported for the first time from off New Caledonia, South Pacific. It is the first study describing the ultrastructure, spermiogenesis and spermatozoon of a species of the family Cavisomatidae. The young spermatid of C. magnum possesses a centriole constituted of doublets without a central element. After the elaboration of a flagellum of 9+2 pattern, the centriole migrates in a nuclear groove. Then the flagellum migration occurs and gives rise to a spermatozoon. The distribution and the size of the protein granules are reported and the posterior extremity appears like a chromatin lamina wave. Comparative ultrastructural data are presented on sperm and spermiogenesis of the Acanthocephala and Rotifers examined to date and the phylogenetic implications are discussed.

Sperm ultrastructure of Helicometra epinepheli (Platyhelminthes, Digenea, Opecoelidae), parasite of Epinephelus fasciatus (Pisces, Teleostei).

This paper describes the ultrastructure of the mature spermatozoon of the digenean Helicometra epinepheli. The ultrastructural elements observed are: two axonemes with a 9+"1" pattern of Trepaxonemata, four attachment zones, a nucleus, two mitochondria, external ornamentation of the plasma membrane, spine-like bodies and cortical microtubules. A particularity of this spermatozoon is the presence of an apical cone. However, the spermatozoon presents the general pattern of the Opecoelidae and thus differs from Helicometra fasciata (which appears as an exception in this family) by several ultrastructural features: the presence of external ornamentation and spine-like bodies and the arrangement of cortical microtubules around the first mitochondrion in the region of the ornamentation. Each ultrastructural feature is discussed and compared with the literature to highlight characters which could be useful for phylogeny.

Spermatozoon ultrastructure of Gyliauchen sp. (Digenea: Gyliauchenidae), an intestinal parasite of Siganus fuscescens (Pisces: Teleostei).

The ultrastructure of the mature spermatozoon of Gyliauchen sp., a parasite of the dusky rabbitfish Siganus fuscescens, was studied by transmission electron microscopy. The spermatozoon possesses two axonemes of the 9+"1" pattern of Trepaxonemata, four attachment zones, one mitochondrion, a nucleus, cortical microtubules, external ornamentation of the plasma membrane, and spine-like bodies. The main characteristics of this spermatozoon are the presence of one mitochondrion, spine-like bodies not associated with the external ornamentation, and a posterior extremity of type 3 that is characterized by the following sequence: posterior extremity of the nucleus then posterior extremity of the second axoneme. Numerous other ultrastructural features are also discussed and compared to the digenean spermatology literature. This is the first study of a member of the Gyliauchenidae and the fourth within the Lepocreadioidea. The results show that many ultrastructural characters are variable within this superfamily and could be useful for phylogeny.

Spermatozoon ultrastructure of Aponurus laguncula (Digenea: Lecithasteridae), a parasite of Aluterus monoceros (Pisces: Teleostei).

The mature spermatozoon of Aponurus laguncula, a parasite of the unicorn leatherjacket Aluterus monoceros, was studied by transmission electron microscopy. The spermatozoon possesses 2 axonemes of the 9+"1" trepaxonematan pattern, attachment zones, a nucleus, a mitochondrion, external ornamentation of the plasma membrane and cortical microtubules. The major features are the presence of: 1) external ornamentation in the anterior part of the spermatozoon not associated with cortical microtubules; 2) one mitochondrion; and 3) cortical microtubules arranged as a single field in the ventral side. The maximum number of microtubules is in the nuclear region. The extremities of the axonemes are characterized by the disappearance of the central core and the presence of microtubule doublets or singlets. This study is the first undertaken with a member of the Lecithasteridae and exemplifies the sperm ultrastructure for the superfamily Hemiuroidea.

Ultrastructural study of the spermatozoon of Heterolebes maculosus (Digenea, Opistholebetidae), a parasite of the porcupinefish Diodon hystrix (Pisces, Teleostei).

This paper describes the ultrastructure of the mature spermatozoon of Heterolebes maculosus. It is the first study of this kind concerning the Opistholebetidae (Platyhelminthes, Digenea). The ultrastructural elements observed in the spermatozoon are: two axonemes with 9+"1" pattern of Trepaxonemata and their attachment zones, two mitochondria, a nucleus, cortical microtubules, external ornamentation of the plasma membrane and spine-like bodies. The number and the disposition of cortical microtubules, the organisation of 11 cortical microtubules disposed in semi-circle around the first mitochondrion in the external ornamentation region and the organisation of the posterior part of the spermatozoon are discussed. Three principal types of posterior part of digenean spermatozoa are proposed. The similarity between the spermatozoon of the Opistholebetidae H. maculosus and Opecoelidae enables us to confirm that these two families are closely related.

Sperm ultrastructure of the gall bladder fluke Anisocoelium capitellatum (Digenea: Cryptogonimidae), a parasite of Uranoscopus scaber (Pisces: Uranoscopidae).

Ultrastructure of mature spermatozoon of Anisocoelium capitellatum is described with transmission electron microscopy. The description gave evidence of some characteristics of this digenean. The male gamete presents features allowing the distinction between A. capitellatum and other digenetic trematodes. It is characterized by original anterior and posterior extremities. The peripheral doublets appear prior to the central cores in the anterior extremity of axoneme. In the distal part of the spermatozoon, these ultrastructural features are also the first elements disappearing in the axoneme 1 and the last remaining in the axoneme 2. Another important peculiarity of the spermatozoon is the presence of three mitochondria.