Vibrio virulence genes in fishes collected from estuarine waters in Italy

Aims: The aim of this study was to investigate the presence of Vibrio vulnificus and potentially pathogenic strains of Vibrio parahaemolyticus in mullets collected from estuarine environment in Italy. Methods and Results: Two hundred and ninety‐five mullets were analysed by culture using the selective medium thiosulfate citrate bile salt sucrose agar, during a monitoring period of 2 years (2008–2009). Presumptive Vibrio colonies were initially identified by using biochemical tests, and strains identified as V. parahaemolyticus and V. vulnificus were subsequently examined by PCR for the presence of species‐specific and virulence genes (toxR, trh, tdh and vvh). V. parahaemolyticus was found in 55% (162/295) of fishes and V. vulnificus in 1% (3/295) with a higher presence in summer months. The trh+/tdh? strains were detected in 16% (47/295) of samples and only one strain resulted trh+/tdh+. One of the V. parahaemolyticus trh+ strains isolated belonged to the O1:KUT (K untypeable), a serotype recently associated to gastroenteritis in Italy. Conclusions: This is the first report demonstrating a high percentage of potential pathogenic V. parahaemolyticus trh+ strains in estuarine fishes of the Mediterranean area. Significance and Impact of the Study: These findings indicate the potential human health risk associated with the presence of pathogenic Vibrio spp. in wild fishes.     

Macrobenthic invertebrate energy densities and ecological status in freshwater watercourses (Friuli Venezia-Giulia,Northeast Italy)

The present study provides energy density (ED) data and models for four macrobenthic invertebrate genera inhabiting freshwater lotic environments (Baetis, Hydropsyche, Rhyacophila, and Onychogomphus). Samples were collected in the hydrological freshwater network of the Region Friuli Venezia Giulia within different watercourse types (creeks, streams, rivers channels, and ditches), and energy density was directly measured using an adiabatic bomb calorimeter. Measured ED expressed in Joule g^?1 wet weight was strongly and positively correlated with percentage of dry weight (DW%) for all genera investigated (r²?>?0.9504), allowing to obtain genus-specific predictive models based on the relationship between ED and DW%. Models were validated and showed good predictive power, as 90th percentile of observed percentage errors ranged between 4.23% and 5.18% while medians ranged between 1.32% and 2.83%. ANCOVA disclosed significant differences between the models, as those for Rhyacophila and Onychogomphus differed significantly from the others. The empirical models were used to build a dataset of estimated energy density, to assess the relationship between energy density and ecological status of the monitored riverine systems, assessed in compliance with European and Italian law by the application of four different ecological indices (ICMi, RQE_IBMR, STAR_ICMi, and LIMeco). Information regarding ED levels for freshwater macrobenthic invertebrates is still neglected in biomonitoring programs, but it could be useful to interpret some ecological situations in the context of ecological status assessment, especially in relation to the trophic condition of the investigated riverine systems.

     

Buccal swab: A tissue sampling method for refinement of experimental procedures involving rainbow trout

Buccal swabbing is a minimally invasive method to obtain DNA and biological material from humans and animals, including fish. Reports on buccal swabbing in fish are few and only for a limited number of species. Rainbow trout (Oncorhynchus mykiss) is an important animal model and because the yield of DNA may vary among and within different species in individuals of different sizes, it was selected as useful to optimize the buccal DNA collection in this species. Different storage methods were evaluated, aimed at DNA preservation by limiting DNA degradation and bacterial growth, using commonly available and inexpensive reagents. DNA quality was also tested by amplification of a single‐copy nuclear gene and a mitochondrial gene. The results suggest that ethanol is the best storage choice for buccal swab sampling in fish genetic studies, as well as suitable for small‐bodied rainbow trout.     

Phosphonate degradation by Spirulina strains: cyanobacterial biofilters for the removal of anticorrosive polyphosphonates from wastewater

The ability of Spirulina spp. to metabolize the recalcitrant xenobiotic Dequest 2054(?) [hexamethylenediamine-N,N,N',N'-tetrakis(methylphosphonic acid)], a CaSO(4) inhibitor used for boiler treatment and reverse osmosis desalination, was investigated. The compound served as sole source of phosphorus, but not of nitrogen, for cyanobacterial growth. In vivo utilization was followed by (31)P NMR analysis. The disappearance of the polyphosphonate proceeded only with actively dividing cells, and no release of inorganic phosphate was evident. However, no difference was found between P-starved and P-fed cultures. Maximal utilization reached 1.0 ± 0.2 mmoll(-1), corresponding to 0.56 ± 0.11 mmol g(-1) dry biomass, thus residual amounts were still present in the exhausted medium when the compound was supplied at higher initial concentrations. At low substrate levels metabolism rates were lower, suggesting that a concentration-driven uptake may represent a limiting step during the biodegradation process. The compound was not retained by biocolumns made with immobilized cyanobacterial cells, either alive or dead. A lab-scale pilot plant, consisting of a series of sequentially connected vessels containing an actively proliferating algal culture, was built and tested for wastewater treatment. Results showed 50% removal of the polyphosphonate added to an initial concentration of 2.5mM. Although further optimization will be required, data strengthen the possibility of using cyanobacterial strains for bioremediation purposes.     

Clonality and Diversity of the Fish Pathogen Lactococcus garvieae in Mediterranean Countries

Infection with Lactococcus garvieae is considered the most important risk factor for the European trout industry, and the losses are approximately 50% of the total production. To improve our understanding of the genetic links among strains originating from different countries, we examined the population structure of L. garvieae by comparing 81 strains isolated from different sources and ecosystems (41 farms in six countries) in which the bacterium is commonly found. Genetic similarities (as assessed with molecular tools, including restriction fragment length polymorphism ribotyping with two endonucleases) were compared with serological data. The combined results reveal that in endemic sites the bacterial population displays a clonal structure, whereas bacterial diversity characterizes sites where the infection is sporadic.     

A survey of bacterial infections in sturgeon farming in Italy

Sturgeon aquaculture production has increased worldwide, particularly in Italy, yet there is little information about diseases that affect sturgeons. With this study we describe the results of screening for bacterial infections during monitoring campaigns carried out from 2014 to 2017. A total of 402 sturgeons from six sturgeon farms with one or more reared sturgeon species among five species and two hybrids were analysed. Bacteriological exam was positive in 93 individuals (23%) and negative in the remaining 309 (77%). Generally, no external or internal macroscopic lesions were present, though some specimens showed skin lesions, liver anaemia or splenomegaly, which are non‐pathognomonic signs of bacterial infection. The majority of the isolated bacteria were opportunistic. This knowledge base is important to help create a bacterial profile of sturgeons reared in Italy.     

Correction to: Changes in midge assemblages (Diptera Chironomidae) in an alpine lake from the Italian Western Alps: the role and importance of fish introduction

Hydrobiologia - Due to an unfortunate turn of events, some given names and family names were transposed in the original publication. The proper representation of the authors’ names and their...     

Occurrence of both subspecies of Photobacterium damselae in mullets collected in the river Magra (Italy)

The natural reservoirs and biological characteristics of pathogenic populations of both subspecies of Photobacterium damselae in aquatic habitants remain unclear because of difficulties in obtaining pathogenic strains from the environment. In the present study, we assessed the occurrence of Photobacterium damselae subsp. piscicida and Photobacterium damselae subsp. damselae , considered to be the causative agent of past epizootic outbreaks in mullets collected in the river Magra, Italy. Two hundred and seventy-eight mullets were collected during a period of two years (2008-2009) and analyzed using multiplex PCR. During this period, 57% of fishes were positive for Photobacterium damselae subsp. piscicida and 37% for Photobacterium damselae subsp. damselae, with an higher presence in summer months although none of PCR-positive mullets showed clinical signs of disease. Our results indicate that the two micro-organisms are widespread in the population of mullets studied, and this could be a possible cause for outbreaks in favourable environmental conditions.     

Seasonal variability of detoxificant response and heavy metal accumulation in tissues of both sexes in <Emphasis Type="Italic">Tinca tinca</Emphasis> (L.) from Lake Trasimeno

Tinca tinca were sampled seasonally from Lake Trasimeno in order to investigate the influence of heavy metal accumulation and changing environmental parameters on the antioxidant responses in this species. Liver, gills and kidney of both sexes were analyzed for total glutathione, catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase, glyoxalase I, glyoxalase II, while muscle was analyzed for arsenic, lead, cadmium and mercury. The MRLs (maximum residue limits) set by actual regulation were not exceeded for heavy metals, but a seasonal and sex-linked variability of antioxidant parameters was observed, and major variations occurred especially in summer and autumn. In this shallow lake, heavy metal accumulation did not affect the biochemical variations, which are rather due to the decreased water level inducing hyperoxia and hypoxia within 24 h, high temperature and loss of mating and refuge areas.     

Enterococcus seriolicida Is a Junior Synonym of Lactococcus garvieae,a Causative Agent of Septicemia and Meningoencephalitis in Fish

The reference strains of Enterococcus seriolicida (ATCC 49156^T) (T = type strain) and of Lactococcus garvieae (ATCC 43921^T) and 30 field strains of Gram-positive cocci isolated from diseased rainbow trout in Italy were found to be phenotypically (API 20 STREPT and API 50 CH) and genetically (DNA-DNA hybridization) similar. The high DNA-DNA homologies (70–100%) and the low ΔT_m(e) (less than 1.1°C) among these strains showed that Enterococcus seriolicida and Lactococcus garvieae are synonyms, describing a single bacterial species. E. seriolicida strains should be classified as L. garvieae, which must be considered as a major pathogen of freshwater and salt water fish with a world-wide distribution.