Hydrocarbons in Seawater and Water Extract of Jarzouna-Bizerte Coastal of Tunisia (Mediterranean Sea): Petroleum Origin Investigation Around Refinery Rejection Place

In this study, the occurrence of toxic heavy metals (As, Cd, Cr, Cu, Pb, and Zn) and relative bioaccumulation in biota samples were investigated in a freshwater ecosystem, the Basento river, one of the main aquatic systems in the south of Italy, which over the last years has been transformed into a sink of urban and industrial wastes. Therefore, the levels of arsenic, cadmium, chromium, copper, lead, and zinc were determined in water, sediments, and tissues of some macroinvertebrate—which are natural assessment endpoints for the evaluation of ecological risk in aquatic systems. Accumulation factors, as a ratio between the concentration of a given contaminant in biota and the one in an abiotic medium, were considered in order to estimate heavy metal contamination loads in biota. Statistical analysis was performed for a comparative evaluation of bioaccumulation among various macroinvertebrates, according to different feeding guilds. The Tukey honestly significantly different test showed significant differences in the bioaccumulation of As, Cd, and Cr among the considered biological receptors (collector–gatherer, predator, and filterer), suggesting that the biological uptake from immediate contact with the sediment or solid substratum (collector–gatherer), instead of the bioconcentration from water (filterer) or biomagnification along the biotic food webs (predators), is the more effective biological sequestering pathway for these metals. Biota–sediment accumulation factors, commonly used for the evaluation of sediment’s role in aquatic systems contamination, were determined for the considered metals. A linear correlation between the concentrations of As, Cd, Cr, and Zn in macroinvertebrates and those in the sediments suggested that the metal uptake data in macroinvertebrates can provide useful information for the estimation of heavy metal exposure risk or bioavailability when making assessments of sediment toxicity in freshwater ecosystems.     

Phylogenetic relationships in grey mullets (Mugilidae) in a Tunisian lagoon

The Mugilidae family is an important fish group representing a major source for fisheries and aquaculture. In the south Mediterranean bank, no data are available on this fauna, except for some morphological studies on Tunisian samples. In this study, 16 allozymic loci were used to investigate the phylogenetic relationships within Tunisian mugilids. The results obtained from Hergla lagoon samples highlight five operational taxonomic unit corresponding to the well‐known species (Liza aurata, Liza ramada, Liza saliens, Chelon labrosus, Mugil cephalus). Several loci appeared to be diagnostic of these species, but in contrast to Greek mugilid samples, we did not find any diagnostic locus fixed differently for the five species. These results can help aquaculture units to identify accurately the mullet species they subsequently use for stocking aquaculture ponds and inland waters. However, species identity represents very important information, as each species has a different growth rate and salinity tolerance. On the other hand, when compared with North Mediterranean Mugilidae analysed until now, Tunisian samples show a genetic differentiation that could be related to different physicochemical conditions between the North and South banks, similar to those recorded in the eastern and western two Mediterranean basins separated by the Siculo‐Tunisian strait. In addition, this study confirms the morphological taxonomy, except for the subdivision of the Liza genus into two sub‐genera. The phylogenetic tree is in agreement with that on Languedoc Mugilidae samples (France), indicating that the subdivision of the Liza genus into two sub‐genera appears to be without any genetic base.     

Ontogeny of the digestive tract of thick lipped grey mullet (Chelon labrosus) larvae reared in “mesocosms”

This work describes the ontogeny of the digestive tract in thick lipped grey mullet (Chelon labrosus) larvae reared until day 36 post-hatching with the semi-extensive technology in mesocosms. Diet was constituted by live preys, rotifers, Artemia and wild zooplankton, then compound diet was added from day 20 (p. h.). Linear growth, weight growth and digestive enzymes specific activities were studied during larval ontogeny. Pancreatic enzymes (trypsin and amylase) and intestinal enzymes (leucine-alanine peptidase “Leu-ala”, aminopeptidase N “AN” and alkaline phosphatase “AP”) were assayed in larvae sampled throughout the rearing trial to evaluate gastrointestinal maturation along the development.The trypsin specific activities were very high during the first two weeks and then declined as observed in marine fish species. A following increase in trypsin specific activity from day 20 was attributed notably to ingestion of particle compound diet. In contrast to the pattern generally described in fish larvae, amylase specific activity showed a continuous increase. This could be attributed to the fact that C. labrosus is an omnivorous species and suggests that the fish might be able to use efficiently diets containing higher levels of starch or other carbohydrates since the end of larval development.Relative expression of intestinal brush border membrane enzymes (AP and AN) and cytosolic enzyme (Leu-ala), showed an abrupt increase of both AP/leu-ala and AN/leu-ala ratios at day 8 (p. h.), indicating that maturation of intestinal tract in C. labrosus larvae is particularly precocious. It is assumed that larvae of C. labrosus might support early co-feeding and weaning strategies, which could reasonably be initiated since mouth opening.     

Melatonin can improve viability and functional integrity of cooled and frozen/thawed rabbit spermatozoa

Melatonin is known to protect sperm against freezing-inflicted damage in different domestic species. The aim of the study was to evaluate the effect of supplementation of semen extender with melatonin on the quality and DNA integrity of cooled and frozen/thawed rabbit spermatozoa. We also investigated whether the addition of melatonin to the semen extender could improve the fertility of rabbit does artificially inseminated with frozen/thawed semen. Semen samples collected from eight rabbit bucks were pooled and then diluted in INRA-82 supplemented either with (0.5, 1.0 or 1.5 mM) or without (0.0 mM) melatonin. Diluted semen was cooled at 5°C for 24 hr. For cryopreservation and based on the first experiment's best result, semen samples were diluted in INRA-82 in the presence or absence of 1.0 mM melatonin and then frozen in 0.25 ml straws. Following cooling or thawing, sperm quality and DNA integrity were evaluated. Furthermore, the fertility of frozen/thawed semen was investigated after artificial insemination. Supplementation of semen extender with 1.0 mM melatonin improved (p < .05) motility, viability, membrane and acrosome integrities in cooled semen compared with other groups. Sperm quality and DNA integrity were higher (p < .05) in frozen/thawed semen diluted in 1.0 mM melatonin-supplemented extender than in the control group. Conception and birth rates were higher in does inseminated with 1.0 mM melatonin treated semen compared with the controls. In conclusion, supplementation of semen extender with 1.0 mM melatonin improved the quality of cooled and frozen/thawed rabbit spermatozoa. Melatonin can preserve DNA integrity and enhance the fertility of frozen/thawed rabbit spermatozoa.     

Effects of biofertilizers and cycocel on some physiological and biochemical traits of wheat (Triticum aestivum L.) under salinity stress

In order to study the effects of biofertilizers and cycocel on some physiological and biochemical characteristics of wheat (Triticum aestivum L.) under salinity condition, a factorial experiment was conducted based on randomized complete block design with three replications under greenhouse condition in 2015. Treatments were included salinity in four levels [no salt (control or S₀), salinity 30 (S₁), 60 (S₂) and 90 (S₃) mM NaCl equivalent of 2.76, 5.53 and 8.3 dS m^?1, respectively], four biofertilizers levels [no biofertilizer (F₀), seed inoculation by Azotobacter chrocoocum strain 5 (F₁), Pseudomonas putida strain 186 (F₂), both inoculation Azotobacter + Pseudomonas (F₃)] and three cycocel levels [without cycocel as control (C₀), application of 600 (C₁) and 1000 (C₂) mg L^?1]. Results showed that salinity severe stress (90 mM) decreased chlorophyll content, relative water content (RWC), total chlorophyll, photochemical efficiency of PSII and yield of wheat. Whereas, soluble sugars and proline content, electrical conductivity (EC), the activity of catalase (CAT), peroxidase (POD), polyphenol oxidase (PPO) enzymes were increased. Similar results were observed in CAT, POD and PPO activities due to inoculation by biofertilizers and cycocel. Salinity at 30 mM increased the photochemical efficiency of PSII and chlorophyll content in plants grown under biofertilizer and cycocel treatment but with increasing salinity up to 90 mM mentioned parameters were decreased. The highest proline and soluble carbohydrate at all salinity levels were observed in plants treated in the highest cycocel level and Azotobacter+ Pseudomonas application. Generally, it was concluded that biofertilizers and cycocel can be used as a proper tool for increasing wheat yield under salinity condition.     

Allometric growth patterns and development in larvae and juveniles of thick‐lipped grey mullet Chelon labrosus reared in mesocosm conditions

Allometric growth and ontogeny were studied in thick‐lipped grey mullet Chelon labrosus reared in mesocosms from 1 to 71 day post hatching (dph). Multivariate allometric analysis of morphometric growth distinguished three distinct developmental stanzas separated by two morphometric metamorphosis lengths (L_m1 = 4.46 ± 0.06 mm; L_m2 = 28.56 ± 1.04 mm). Body mass growth also showed three distinct episodes separated by two inflections, correlated with morpho‐functional changes. First episode concerned pre‐flexion larvae and ended around 4.5 mm‐L_T (14‐dph), coinciding with estimated L_m1. It was distinguished by reduced growth, but intense morphogenesis and differentiation processes. Organogenesis and allometric changes indicated that development priorities concerned feeding efficiency, by improving detection ability (sensory system development), ingestion capacity (head growth) and assimilation performance (digestive system differentiation), together with respiration efficiency (gill development). Second episode concerned post‐flexion larvae and, ended around 8.6 mm‐L_T (25‐dph). It was distinguished by fast growth of trunk and tail, acquisition of adult axial muscle distribution and completion of gill filament development, improving locomotion and oxygenation performances. It corresponded to transition towards metamorphosing stage as indicated by later isometric growth, musculature maturation and acquisition of juvenile phenotype. Metamorphosis seemed to end at L_m2, suggesting to avoid zootechnic handling before this size.     

Experimental approach on the selection and persistence of anti-microbial-resistant Aeromonads in faecal matter of rainbow trout during and after an oxolinic acid treatment

The presence of anti-microbial-resistant bacteria in the aquatic environment remains a matter of concern for environmental, animal and human health risks. Many published data have reported increased anti-microbial resistance among the aquatic microbiota from fish farm environments which often coincides with the use of anti-microbial agents. It can be assumed that the selection of anti-microbial-resistant bacteria first occurs in the fish farms themselves, at locations where anti-microbial concentrations are active. As anti-microbial agents are frequently administered through feed pellets, faecal matter excreted could favour conditions for such selection. In this study, OA (oxolinic acid) concentrations and OA resistance of Aeromonads were surveyed in output water and in faecal matter from OA medicated (test) and non-medicated (control) fish tanks during and after an OA oral treatment. Because of the low counts of Aeromonads in the output water, it was not possible to obtain reliable data on the proportion of anti-microbial resistance in this compartment. On the other hand, a time-limited effect on the abundance of Aeromonads isolated from faecal matter was only observed in test tanks. In these tanks, high proportion (80-100%) of OA-resistant Aeromonads was present in deposited faecal matter until at least one week after the end of the anti-microbial treatment. At that time OA was no longer detected within the faecal matter, after having reached concentrations between 100 and 190 μg/g during the treatment. We discuss how the leaching of OA from faecal pellets, from the intestine to their deposit, may favour the selection of OA-resistant Aeromonads. Further investigations in real conditions on microbiota associated with faecal matter in transit through the fish farm system are required. Particular attention should be paid to the wastes collected by the filter treatments of fish farm effluents.     

基于黄河鲤新品系IGF2b基因SNPs获取体系的建立及在该品系选育中的应用

为了建立鲤 IGF2b (insulin-like growth factor 2)基因全基因组信息 SNPs (single nucleotide polymorphism)的获取体系,并验证该获取体系在黄河鲤新品系选育中运用的适用性,本研究首先对 10个不同鲤品种的 33个鲤个体重测序数据进行分析,整体上了解 IGF2b 基因的单核苷酸位点及其在基因组上的分布情况。然后在基因组 DNA 水平上分段获取 IGF2b 基因的 SNPs 信息,并以不同的鲤品种验证引物的适用性,最后在黄河鲤新品系中检测其应用。研究结果:获得 8 对聚合酶链式反应(polymerase chain reaction, PCR)引物进行分段克隆的结果,并通过直接测序和限制性片段长度多态性来获取相应的 SNPs。经过直接测序,本文对黄河鲤、建鲤、黄河鲤和建鲤的正反交后代进行 PCR 检测发现条带单一,目地片段正确;结合黄河鲤新品系的体重数据,本文又以 IGF2b3#引物检测到一个与体重明显相关的 SNPs,同时检测到一个与该基因表达量相关的另一个 SNPs。本研究的方法能够在该基因全基因组范围内开展鲤分子育种中 SNPs 的检测,并验证发现黄河鲤新品系 IGF2b3#引物 227 号位点处,出现的突变位点使体重降低,且有一处 SNPs 与其表达量有关。这将为其他物种、其他功能基因的多态性分子标记研究提供思路。