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Aquaculture farms in Arkansas, USA routinely battle columnaris disease caused by Flavobacterium covae. Columnaris is prevalent during stressful events such as feed training and when fish are stocked at high densities in holding vats before sale. Kaolin clay was effective in laboratory trials as a treatment for columnaris in catfish. As a result, fish farmers are interested in applying kaolin products but were hesitant as they feared that the high doses of kaolin clay in vats might negatively affect the gills and overall health of fish. Therefore, we evaluated potential clay concentrations that might be used to prophylactically treat fish in vats. The effects of low to excessively high doses (0, 1, 2, 4, or 8 g/L) of kaolin clay (AkuaProTM, Imerys, GA, USA) were evaluated using a 72 h bioassay conducted in static tanks using Micropterus salmoides, Pomoxis nigromaculatus, Lepomis macrochirus, Ictalurus punctatus, Notemigonus crysoleucas, and Pimephales promelas. Results of these trials revealed a 100% survival rate across all six fish species exposed to kaolin clay at concentrations of up to 8 g/L for 48 h (followed by a 24 h recovery period in clean water) with no adverse effects to eyes, skin, gastrointestinal tract, or liver histology noted at any treatment. In addition, Micropterus salmoides analyzed for heavy metals due to exposure to the clay indicated that concentrations did not differ from control fish.

The present study was aimed to explore the phytochemical composition, antioxidant potential, and density function theory (DFT) study of Araucaria columnaris (G. Forst.) Hook. leaves. Different extraction techniques like Soxhlet, ultrasound, and maceration were performed using solvents—hexane, chloroform, and methanol. Among the three solvents, the best antioxidant values were observed in methanolic extracts followed by chloroform and hexane. The results of RP-HPLC-DAD showed the 11 polyphenolic compounds in methanolic extracts. The observed order of yield, polyphenolic content and antioxidant potential in three techniques was SOX > ULT > MCE. For the first time, 48 bioactive compounds were identified in methanol, chloroform, and hexane extracts using GC–MS technique. DFT has been carried out on major phytochemicals of methanol extract at B3LYP/6–31G (d) basis set (Gaussian 09 program). The obtained results concluded that the A. columnaris (G. Forst.) Hook. leaves contained the distinct phytochemicals having ethnopharmacological employments and significant antioxidant potential.

Columnaris disease generates substantial losses of many freshwater fish species; one is the hybrid striped bass. The ubiquitous aquatic bacterium Flavobacterium columnare can be highly effective in biofilm formation on fish skin and gills. Previous research showed a difference between columnaris disease susceptibility of hybrid striped bass (Morone saxatilis × M. chrysops) and white bass (M. chrysops). To understand these differential susceptibilities and possible mucosal relationship, we assessed total bacterial growth and biofilm formation with mucus derived from each moronid parental species: white bass and striped bass (M. saxatilis). Differential susceptibility was confirmed of the other parent species, the striped bass (M. saxatilis). In addition to intraspecies investigations, individual hybrid striped bass mucosal affects were also studied for deferential responses to bacterial growth and biofilm formation. Species- and concentration-dependent differences were detected in the total growth of the bacteria to host mucus. Our data suggest that bass mucus can significantly affect biofilm formation with the F. columnare isolate tested. There appears to be a correlation between the bacteria's response of growth and biofilms and bass species susceptibility. This study provides insight into our understanding of the host-pathogen interaction between F. columnare and moronids.

The article presents the results of studies on the treatment scheme efficacy for columnaris in Pterophyllum scalare, common under private aquarium husbandry conditions. To establish the diagnosis, the clinical features of the diseased fish, pathological and anatomical changes and the results of microscopic and microbiological studies were taken into account. Separate chemical and microbiological parameters of aquarium water were also studied. It was established that fish disease developed against the background of adverse changes in the chemical composition and microbiocenosis of aquarium water. High alkalinity and excess of phosphates compared to the norm provoked accumulation of opportunistic microbiota, resulting in a balance disorder in the parasite-host system and development of clinical manifestation of the fish disease. During the disease outbreak, bacteriological indices of water indicated a high level of organic contamination and a low intensity of water self-purification processes. Clinically, the disease was manifested in P. scalare by decrease in appetite and motor activity, onset of ulcerative lesions of various shapes and sizes on the surface of the body and on the gill covers. Selected pure cultures of Flavobacterium columnare showed sensitivity to enrofloxacin (growth retardation zone 31.3 ± 1.0 mm); moderate resistance was found to tylosin. The microorganisms were resistant to amoxicillin, doxycycline, benzylpenicillin and tetracycline. Microscopic studies of intestinal specimens of dead P. scalare revealed numerous motile flagellates. It has been shown that an effective treatment regimen that provides recovery for 70% of diseased P. scalare is the use of enroxil 10% solution for five days, metronidazole three times a day, and “API MelaFix” for seven days. It is proved that the following measures are effective to restore the disrupted hydro-balance: periodic water replacement in the amount of 20% of the total volume, providing the aquarium with active aeration systems, planting slow-growing plants and reducing the amount of fish food provided. The measures developed were efficient, they led to elimination of the outbreak of columnaris in the P. scalare and to restoration of biological equilibrium in a closed aquatic ecosystem.

Catfish is an important aquaculture species in the USA. Columnaris disease is distributed worldwide, affecting a wide variety of fish species including catfish . It leads to huge economic losses each year to the US catfish industry. Channel catfish in general is highly resistant to the disease, while blue catfish is highly susceptible. Genomic selection is an effective and accurate way to predict the breeding values and thus was expected to improve the prediction veracity of columnaris disease resistance in catfish effectively. In this study, two different methods, elastic net genomic best linear unbiased prediction (ENGBLUP) and genomic best linear unbiased prediction (GBLUP), were used to predict the columnaris disease resistance evaluated by binary survival status. Cross-validation showed that the prediction accuracy of ENGBLUP and GBLUP was 0.7347 and 0.4868, respectively, showing that ENGBLUP had a high prediction accuracy. It was shown that fitting QTL and polygenic effect with different distribution will improve genomic prediction accuracy for binary traits. In this study, an accurate and effective genomic selection method was proposed to predict the columnaris resistance in catfish, and its application should be beneficial to catfish breeding.

Abstract This work investigates the removal of phenol from aqueous solution using Araucaria Columnaris bark (ACB) as biochar. Five different types of biochars were developed through pyrolysis at different temp from 300 to 500°C. The effects of initial concentration, contact time, pH and temperature on adsorption behavior were studied in batch mode for each biochar. The optimum contact time observed for equilibrium condition was 60 mins for every biochar. And, the maximum adsorption followed the order 298 K > 308 K > 318 K. Adsorption equilibrium data were fitted to Langmuir and Freundlich isotherms by non-linear regression method and kinetic data by linear regression method, and fitted to pseudo-first order, pseudo-second order and Intraparticle diffusion models. Adsorption kinetics was reasonably described by pseudo-second order model with R 2 value 0.99. Thermodynamic parameters were also estimated that implied, the adsorption process was spontaneous and exothermic in nature. Study further showed that the acidic pH increased adsorption capacity of biochar but decreases continuously towards basic side. The removal of phenol with prepared biochar was achieved as high as 100 % for ACB-500. The maximum iodine adsorption value of prepared biochar was found to be 453.3 mg/g.

In the present study, phytochemical investigation, antioxidant potential and quantum study on the dichloromethane (DCM) and ethanol extracts of Araucaria columnaris (G. Forst.) Hook. branches have been carried out. The ethanol extract has higher total bioactive contents as compare with DCM extract such as total polyphenolic contents, total flavonoids contents and total tannin contents. The phytochemical quantification and volatile composition of most bioactive extract (ethanol) has been carried out by RP-HPLC-DAD (9-compounds) and GC-MS (19-compounds). The antioxidant potential was demonstrated using free radical scavenging, phosphomolybdenum, metal chelating and reducing power assays. The ethanol extract has higher antioxidant potential. The Density Functional Theory (DFT) has been carried out on major phytochemicals of ethanol extract. The current findings revealed an excellent concentration of bioactive compounds in A. columnaris (G. Forst.) Hook. branch, which reaffirms the great significance of the current work and benefaction to natural products chemistry.

Background Columnaris disease (CD) is an emerging problem for the rainbow trout aquaculture industry in the US. The objectives of this study were to: (1) identify common genomic regions that explain a large proportion of the additive genetic variance for resistance to CD in two rainbow trout (Oncorhynchus mykiss) populations; and (2) estimate the gains in prediction accuracy when genomic information is used to evaluate the genetic potential of survival to columnaris infection in each population. Methods Two aquaculture populations were investigated: the National Center for Cool and Cold Water Aquaculture (NCCCWA) odd-year line and the Troutlodge, Inc., May odd-year (TLUM) nucleus breeding population. Fish that survived to 21 days post-immersion challenge were recorded as resistant. Single nucleotide polymorphism (SNP) genotypes were available for 1185 and 1137 fish from NCCCWA and TLUM, respectively. SNP effects and variances were estimated using the weighted single-step genomic best linear unbiased prediction (BLUP) for genome-wide association. Genomic regions that explained more than 1% of the additive genetic variance were considered to be associated with resistance to CD. Predictive ability was calculated in a fivefold cross-validation scheme and using a linear regression method. Results Validation on adjusted phenotypes provided a prediction accuracy close to zero, due to the binary nature of the trait. Using breeding values computed from the complete data as benchmark improved prediction accuracy of genomic models by about 40% compared to the pedigree-based BLUP. Fourteen windows located on six chromosomes were associated with resistance to CD in the NCCCWA population, of which two windows on chromosome Omy 17 jointly explained more than 10% of the additive genetic variance. Twenty-six windows located on 13 chromosomes were associated with resistance to CD in the TLUM population. Only four associated genomic regions overlapped with quantitative trait loci (QTL) between both populations. Conclusions Our results suggest that genome-wide selection for resistance to CD in rainbow trout has greater potential than selection for a few target genomic regions that were found to be associated to resistance to CD due to the polygenic architecture of this trait, and because the QTL associated with resistance to CD are not sufficiently informative for selection decisions across populations. Electronic supplementary material The online version of this article (10.1186/s12711-019-0484-4) contains supplementary material, which is available to authorized users.

Columnaris is one of the oldest known fish diseases caused by Flavobacterium columnare. The wide spreading of the causative agent in freshwater environments and the susceptibility of fish to be attacked by it after mechanical and/or environmental factors makes F. columnare among the most prevalent pathogens in cultured, ornamental, and wild fish populations. One hundred fishes of tilapia (Oreochromis niloticus) were collected randomly from ponds of Central Laboratory for Aquaculture Research of Abbassa, Abou Hammad, Sharkia, Egypt for Flavobacterium columnare isolation. F. columnare was isolated from (150) out of the (350) examined samples. The bacteriological examination revealed that the prevalence of F. columnare was (42.8%) in Oreochromis niloticus. The highest prevalence of F columnare was in gills (33.3%) followed by skin (30%) then fins (24.6%) and kidney (12%). These isolates were further characterized by PCR that revealed positive result for 16S RNA gene with specific amplicons size 675bp. The result of antimicrobial sensitivity testing of the isolates revealed that doxycycline, erythromycin, tetracycline, trimethoprim, nalidixic acid and streptomycin were the most effective drugs against F.columnare.

We applied fluorescent in situ hybridization (FISH) using (GAA)10 microsatellite repeat alone or in combination with oligo pSc119.2-1, pTa535-1, pAs1-1, 5S and 45S rDNA probes to identify individual chromosomes and produce banded karyotypes for five allopolyploid Aegilops species that were thought to possess M subgenome. Labelling patterns of GAA repeats allow identification of individual chromosomes in the studied species including diploid Ae. comosa subsp. heldreichii (2n = MhMh) and Ae. umbellulata (2n = UU); and allopolyploid Ae. biuncialis (2n = UbUbMbMb), Ae. geniculata (2n = UgUgMgMg), Ae. columnaris (2n = UcUcXcXc), Ae. neglecta (2n = UnUnXnXn) and Ae. crassa (2n = 4x = D1D1XcrXcr and 2n = 6x = D1D1XcrXcrDcrDcr). Intraspecific polymorphisms of GAA labelling patterns were observed between different genotypes in each species. FISH results showed that the X* subgenomes of Ae. columnaris and Ae. neglecta are highly similar, implying that both species originated from a common ancestor. However, these X* subgenomes were also distinct from and more heterochromatic when compared to the M* subgenomes of Ae. biuncialis and Ae. geniculata. This suggests that the X* subgenomes may not have originated from the Ae. comosa, as previously thought. Although (GAA)n bands are relatively few and weak in Ae. tauschii (DD genome), all 4x Ae. crassa (D1D1XcrXcr genome) chromosomes showed distinct GAA patterns. (GAA)10 probe alone was not sufficient for the discrimination of all the chromosomes of 6x Ae. crassa (D1D1XcrXcrDcrDcr), which needs a combination of (GAA)10 with the pTa535-1 or pAs1-1 probes.

Gram-negative bacteria is the main causative agents for columnaris disease outbreak to finfishes. The outer membrane proteins (OMPs) candidate of Flavobacterium columnare bacterial cell served a cr...

Antimicrobial resistance is a global concern, especially with the backdrop of the development of possible environmental and public health hazards. Flavobacterium columnare has been observed to be multi-drug resistant but highly susceptible to sodium chloride, hence the need for susceptibility profiling. Therapeutic efficacy of commonly used antibiotics and sodium chloride was tested in-vitro against four isolates of F. columnare using the disc diffusion and the pour plate methods, respectively. Comparative in-vivo testing was performed on experimentally infected Clarias gariepinus juveniles in 11 groups of 15 fish each. Nine groups were treated with 1.0, 2.0 and 3.0% NaCl for 5, 10 and 30 min each. A tenth group was exposed to 25 mg L-1 ciprofloxacin for 1 h by immersion, while the eleventh group was not treated (positive control). Growth inhibition was observed to be highest with ciprofloxacin followed by ofloxacin and tetracycline in that order, and also in all NaCl concentrations. There was no mortality in the infected fish groups treated with 25 mg L-1 ciprofloxacin, 1% NaCl, and 2 to 3% NaCl for 5 and 10 min only. However, 46.7 ± 9.4 % mortality observed in 3 % NaCl treatment for 30 min was significantly higher than the 23.3 ± 4.6% observed in 2 % NaCl for 30 min, but not significantly different from the positive control with 36.7±4.7 % mortality. Since short duration sodium chloride bath was found to be effective in the control of columnaris disease, 1% salt disinfection of fry and fingerling stock for 30 min could be incorporated into routine management in catfish hatchery, without the fear of environmental or public health hazards. Key words: Antibiogram, columnaris disease, sodium chloride, susceptibility.

Vaccination is the most effective approach for prevention of infectious diseases in aquaculture. Although immersion vaccination is more applicable compared to in-feed/oral administration and injection, this method suffers from low potency as the efficiency of uptake of antigens through mucosal membranes is limited. In this study, we have successfully developed a mucoadhesive vaccine delivery system to enhance the efficacy of direct immersion vaccination against Flavobacterium columnare, the causative agent of columnaris disease in red tilapia. A formalin-killed negatively charged, bacterial cell suspension was used to prepare a mucoadhesive vaccine by electrostatic coating with positively charged chitosan. Our results demonstrate that the chitosan-complexed vaccine greatly increases its mucoadhesiveness, thus increasing the chances of vaccine uptake by the gill mucosa and improving the protection obtained against columnaris infection. The surface charge of the chitosan-complexed vaccine was altered from anionic to cationic after chitosan modification. Tilapia were vaccinated with the prepared chitosan-complexed vaccine by immersion. The challenge test was then carried out 30 and 60 days post vaccination, which resulted in a high level of mortalities in the non-vaccinated and uncomplexed vaccine groups. A high relative percentage survival (RPS) of vaccinated fish was noted with the mucoadhesive vaccine. Our results indicated that the naked vaccine failed to protect the fish from columnaris infection, which is consistent with the mucoadhesive assays performed during the study showing that the naked vaccine was unable to bind to mucosal surfaces. This system is therefore an effective method for immersion vaccination in order to deliver the antigen preparation to the mucosal surface membrane of the fish.

Flavobacterium columnare is the causative agent of columnaris disease. Previous work has demonstrated a high degree of genetic variability among F. columnare isolates, identifying 4 genetic groups (GGs) with some host associations. Herein, a total of 49 F. columnare isolates were characterized, the majority of which were collected from 15 different locations throughout the US Pacific Northwest. Most isolates were collected from 2015-2018 and originated from disease outbreaks in salmonid hatcheries and rearing ponds, sturgeon hatcheries and ornamental fish. Other isolates were part of collections recovered from 1980-2018. Initial identification was confirmed by F. columnare species-specific qPCR. Study isolates were further characterized using a multiplex PCR that differentiates between the 4 currently recognized F. columnare GGs. Multiplex PCR results were supported by repetitive sequence-mediated PCR fingerprinting and gyrB sequence analysis. F. columnare GG1 was the most prevalent (83.7%, n = 41/49), represented by isolates from salmonids (n = 32), white sturgeon (n = 2), channel catfish (n = 1), ornamental goldfish (n = 1), koi (n = 3), wild sunfish (n = 1) and 1 unknown host. Six isolates (12.2%, n = 6/49) were identified as GG3, which were cultured from rainbow trout (n = 3) and steelhead trout (n = 3). Two isolates were identified as GG2 (4.1%, n = 2/49) and were from ornamental fish. No GG4 isolates were cultured in this study. The biological significance of this genetic variability remains unclear, but this variation could have significant implications for fish health management. The results from this study provide baseline data for future work developing strategies to ameliorate columnaris-related losses in the US Pacific Northwest.

The present study aimed to assess the efficiency of silver bio-nanoparticles (Ag-NPs) in inactivating of the Aspergillus fumigatus, A. parasiticus and A. flavus var. columnaris and A. aculeatus spores. The AgNPs were synthesized in secondary metabolic products of Penicillium pedernalens 604 EAN. The inactivation process was optimized by response surface methodology (RSM) as a function of Ag NPs volume (1–10 μL/mL); time (10–120 min); pH (5–8); initial fungal concentrations (log10) (3–6). The artificial neural network (ANN) model was used to understand the behavior of spores for the factors affecting inactivation process. The best conditions to achieved SAL 10−6 of the fungal spores were recorded with 3.46 μl/mL of AgNPs, after 120 min at pH 5 and with 6 log of initial fungal spore concentrations, at which 5.99 vs. 6.09 (SAL 10−6) log reduction was recorded in actual and predicted results respectively with coefficient of 87.00%. The ANN revealed that the timehas major contribution in the inactivation process compare to Ag NPs volume. The fungal spores were totally inactivated (SAL 10−6, 6 log reduction with 99.9999%) after 110 min of the inactivation process, 10 min more was required to insure the irreversible inactivation of the fungal spores. The absence of protease and cellulase enzymes production confirm the total inactivation of the fungal spores. FESEM analysis revealed that the AgNPs which penetrated the fungal spores leading to damage and deform the fungal spore morphology. The AFM analysis confirmed the total spore surface damage. The bands in the range of the Raman spectroscopy from 1300 to 1600 cm−1 in the inactivated spores indicate the presence of CH3, CH2 and the deformation of lipids released outside the spore cytoplasm. These finding indicate that the AgNPs has high potential as a green alternative inactivation process for the airborne fungal spores.

We aimed to infer the phylogenetic relationships of populations of Lobelia columnaris using chloroplast genomes and estimate the divergence time to reconstruct its historical colonization on the sky islands of Bioko and Cameroon. Specifically, we aim to answer the following questions: (1) What is the phylogenetic relationship among Bioko Island and Cameroon populations? (2) Are the older populations found on the older sky islands? (3) Does the colonization history reflect the age of the sky islands? We assembled novel plastomes from 20 individuals of L. columnaris from five mountain systems. The plastome data was explored with phylogenetic analyses using Maximum likelihood and Bayesian Inference. The complete plastome size varied from 164,609 bp to 165,368 bp. The populations of L. columnaris have a monophyletic origin, subdivided into three plastome-geographic clades. The plastid phylogenomic results and age of the sky islands indicate that L. columnaris colonized first along the Cameroon Volcanic Line’s young sky islands. The earliest divergent event (1.54 Ma) split the population in South Bioko from those on the mainland and North Bioko. The population of South Bioko was likely isolated during cold and dry conditions in forest refugia. Presumably, the colonization history occurred during the middle-late Pleistocene from South Bioko’s young sky island to North Bioko and the northern old sky islands in Cameroon. Furthermore, the central depression with lowland forest between North and South Bioko is a current geographic barrier that keeps separate the populations of Bioko from each other and the mainland populations. The Pleistocene climatic oscillations led to the divergence of the Cameroon and Bioko populations into three clades. L. columnaris colonized the older sky island in mainland Cameroon after establishing South Bioko’s younger sky islands. The biogeography history was an inverse progression concerning the age of the Afromontane sky islands.

Thirty wheat flour samples were collected from different bakeries and markets in Qena city, Egypt over five months were used to measure moisture content, fungi and aflatoxins. Moisture contents ranged from 5.92 % to 14.43%. Twenty-one fungal species belonging to 13 genera were isolated from the wheat flour samples on Czapeks agar media at 28 ᵒC. The most common fungal genera were Aspergillus, Mucor and Penicillium. The correlation coefficient analysis revealed to strong positive correlation between moisture content and average total count of fungi in wheat flour samples (0.92). Qualitative and quantitative determination of aflatoxins in 29 isolates of the A. flavi group (A. flavus, A. flavus var. columnaris and A. parasiticus) were made using thin layer chromatography (TLC) and high performance liquid chromatography (HPLC), respectively. The results for (TLC) were as follow: fourteen isolates (48.2%) had the ability to produce aflatoxin B1, five isolates (17.2%) could produce both aflatoxin B1 and aflatoxin B2 and ten (34.4%) isolates were negative. By using HPLC, the concentrations of aflatoxin were 13.416–9229.343, 2.639–152.668 and 6.391–79.507µg/L for AFB1, AFB2, and AFG2, respectively.

Columnaris, a highly contagious bacterial disease caused by Flavobacterium columnare, is recognized as one of the most important infectious diseases in farmed tilapia, especially during the fry and fingerling stages of production. The disease is associated with characteristic lesions in the mucosa of affected fish, particularly their skin and gills. Vaccines delivered via the mucosa are therefore of great interest to scientists developing vaccines for this disease. In the present study, we characterized field isolates of F. columnare obtained from clinical columnaris outbreaks in red tilapia to select an isolate to use as a candidate for our vaccine study. This included characterizing its colony morphology, genotype and virulence status. The isolate was incorporated into a mucoadhesive polymer chitosan-complexed nanovaccine (CS-NE), the efficacy of which was determined by experimentally infecting red tilapia that had been vaccinated with the nanoparticles by immersion. The experimental infection was performed 30-days post-vaccination (dpv), which resulted in 89% of the unvaccinated control fish dying, while the relative percentage survival (RPS) of the CS-NE vaccinated group was 78%. Histology of the mucosal associated lymphoid tissue (MALT) showed a significantly higher presence of leucocytes and a greater antigen uptake by the mucosal epithelium in CS-NE vaccinated fish compared to control fish and whole cell vaccinated fish, respectively, and there was statistically significant up-regulation of IgT, IgM, TNF α, IL1-β and MHC-1 genes in the gill of the CS-NE vaccinated group. Overall, the results of our study confirmed that the CS-NE particles achieved better adsorption onto the mucosal surfaces of the fish, elicited great vaccine efficacy and modulated the MALT immune response better than the conventional whole cell-killed vaccine, demonstrating the feasibility of the mucoadhesive nano-immersion vaccine as an effective delivery system for the induction of a mucosal immune response against columnaris disease in tilapia.

The role of endophytic fungi isolated from different populations of European Ni hyperaccumulators was investigated in regard to the microorganisms' ability to enhance the hyperaccumulation of Ni in Noccaea caerulescens. Effects of particular species of endophytic fungi on adaptation of N. caerulescens to excess Ni were tested by co-cultivation with single strains of the fungi. Seven of these had a positive effect on plant biomass production, whereas two of the tested species inhibited plant growth; biomass production of inoculated plants was significantly different compared to non-inoculated control. Inoculation with six fungal strains: Embellisia thlaspis, Pyrenochaeta cava, Phomopsis columnaris, Plectosphaerella cucumerina, Cladosporium cladosporioides and Alternaria sp. stimulated the plant to uptake and accumulate more Ni in both roots and shoots, compared to non-inoculated control. P. columnaris was isolated from all plant species sampled. Strains isolated from Noccaea caerulescens and Noccaea goesingensis increased Ni root and shoot accumulation of their native hosts (compared to non-inoculated control). Inoculation of different populations of Noccaea with P. columnaris of foreign origin did not cause its host to accumulate more Ni, with the exception of the Ni-unadapted ecotype of N. goesingensis. Inoculation with P. columnaris from N. caerulescens significantly improved Ni uptake, but the effect of the fungus was not as prominent as in the case of N. caerulescens. By comparing the transcriptomes of N. caerulescens and N. goesingensis from Flatz inoculated with P. columnaris, we showed that enhanced uptake and accumulation of Ni in the plants is accompanied by an upregulation of several genes mainly involved in plant stress protection and metal uptake and compartmentation.

Phenotypic variation is suggested to facilitate the persistence of environmentally growing pathogens under environmental change. Here, we hypothesized that the intensive farming environment induces higher phenotypic variation in microbial pathogens than natural environment, because of high stochasticity for growth and stronger survival selection compared to the natural environment. We tested the hypothesis with an opportunistic fish pathogen Flavobacterium columnare isolated either from fish farms or from natural waters. We measured growth parameters of two morphotypes from all isolates in different resource concentrations and two temperatures relevant for the occurrence of disease epidemics at farms and tested their virulence using a zebrafish (Danio rerio) infection model. According to our hypothesis, isolates originating from the fish farms had higher phenotypic variation in growth between the morphotypes than the isolates from natural waters. The difference was more pronounced in higher resource concentrations and the higher temperature, suggesting that phenotypic variation is driven by the exploitation of increased outside-host resources at farms. Phenotypic variation of virulence was not observed based on isolate origin but only based on morphotype. However, when in contact with the larger fish, the less virulent morphotype of some of the isolates also had high virulence. As the less virulent morphotype also had higher growth rate in outside-host resources, the results suggest that both morphotypes can contribute to F. columnare epidemics at fish farms, especially with current prospects of warming temperatures. Our results suggest that higher phenotypic variation per se does not lead to higher virulence, but that environmental conditions at fish farms could select isolates with high phenotypic variation in bacterial population and hence affect evolution in F. columnare at fish farms. Our results highlight the multifaceted effects of human-induced environmental alterations in shaping epidemiology and evolution in microbial pathogens.

This study evaluates the effects of dietary inclusion of grape pomace flour (GPF) on growth, antioxidant, anti-inflammatory, innate-adaptive immunity, and immune genes expression in Labeo rohita against Flavobacterium columnaris. In both normal and challenged fish the growth rate, hematology and biochemical parameters significantly increased when fed with 200 and 300 mg GPF enriched diets; similarly the activities of antioxidants and innate-adaptive immune parameters, such as malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione (GSH), phagocytic (PC), respiratory burst (RB), alternative pathway complement (ACP), lysozyme (Lyz), and total immunoglobulin M (IgM) significantly increased in both groups. Similarly, the immune, antioxidant, and anti-inflammatory-related gene mRNA expression was significantly up-regulated in head kidney (HK) tissues. The challenged fish fed without GPF always exhibited lower values of all the studied parameters. The results indicate that both normal and challenged fish treated with 200 mg GPF inclusion diet had significantly enhanced growth rate, antioxidant status, and immune defense mechanisms than with 300 mg GPF diet in L. rohita against F. columnaris.

Fish diseases are the major cause for increasing mortality in fish farms. Automatic identification of diseased fish at early stages is necessary step to prevent spreading disease. Fish disease diagnosis suffers from some limitations that need high level of expertise to be resolved. Recognition of fish abnormal behaviors helps in early prediction of fish diseases. Fish behavior is evaluated by analyzing fish trajectories in videos. Abnormalities may be due to environmental changes. This paper introduces a survey on what computer vision techniques propose in that field. A comprehensive comparison between different automatic recognition systems is included. Finally, our approach is proposed to automatically recognize and identify three different types of fish diseases. These diseases are Epizootic ulcerative syndrome (EUS), Ichthyophthirius (Ich) and Columnaris. Our approach shows the effect of different color spaces on the Convolutional Neural Networkk CNN final performance.

Background Drug-resistant fish pathogens can cause significant economic loss to fish farmers. Since 2012, florfenicol has become an approved drug for treating both septicemia and columnaris diseases in freshwater fish. Due to the limited drug options available for aquaculture, the impact of the therapeutical florfenicol treatment on the microbiota landscape as well as the resistome present in the aquaculture farm environment needs to be evaluated. Results Time-series metagenomic analyses were conducted to the aquatic microbiota present in the tank-based catfish production systems, in which catfish received standard therapeutic 10-day florfenicol treatment following the federal veterinary regulations. Results showed that the florfenicol treatment shifted the structure of the microbiota and reduced the biodiversity of it by acting as a strong stressor. Planctomycetes, Chloroflexi, and 13 other phyla were susceptible to the florfenicol treatment and their abundance was inhibited by the treatment. In contrast, the abundance of several bacteria belonging to the Proteobacteria, Bacteroidetes, Actinobacteria, and Verrucomicrobia phyla increased. These bacteria with increased abundance either harbor florfenicol-resistant genes (FRGs) or had beneficial mutations. The florfenicol treatment promoted the proliferation of florfenicol-resistant genes. The copy number of phenicol-specific resistance genes as well as multiple classes of antibiotic-resistant genes (ARGs) exhibited strong correlations across different genetic exchange communities (p Conclusions The therapeutical level of florfenicol treatment significantly altered the microbiome and resistome present in catfish tanks. Both intra-population and inter-population horizontal ARG transfer was observed, with the intra-population transfer being more common. The oxazolidinone/phenicol-resistant gene optrA was the most prevalent transferred ARG. In addition to horizontal gene transfer, bacteria could also acquire florfenicol resistance by regulating the innate efflux systems via mutations. The observations made by this study are of great importance for guiding the strategic use of florfenicol, thus preventing the formation, persistence, and spreading of florfenicol-resistant bacteria and resistance genes in aquaculture.

Aegilops columnaris is a tetraploid species originated from Ae. umbellulata (2n=2x=14, UU) and a yet unknown diploid grass species. Although Ae. columnaris possesses some agronomically valuable traits, such as heat and drought tolerance and resistance to pests, it has never been used in wheat breeding because of difficulties in producing hybrids and a lack of information on the relationships between Ae. columnaris and common wheat chromosomes. In this paper, we report the development of 57 wheat – Ae. columnaris introgressive lines covering 8 of the14 chromosomes of Aegilops. Based on substitution spectra of hybrids and the results of FISH analysis of the parental Ae. columnaris line with seven DNA probes, we have developed the genetic nomenclature of the Uc and Xc chromosomes. Genetic groups and genome affinities were established for 11 of 14 chromosomes; the classification of the remaining three chromosomes remains unsolved. Each Ae. columnaris chromosome was characterized on the basis of C-banding pattern and the distribution of seven DNA sequences. Introgression processes were shown to depend on the parental wheat genotype and the level of divergence of homoeologous chromosomes. We found that lines carrying chromosome 5Xc are resistant to leaf rust; therefore, this chromosome could possess novel resistance genes that have never been utilized in wheat breeding.

The reconstruction of past environments has been studied in several areas of science, aiming to help in understanding the several events that occurred on the planet. Plants are considered environmental markers and usually show structural adaptations that are characteristic of their habitat. Therefore, after passing through the charring process, they may not be completely consumed by the fire, being possible to find remnants of charcoal and other remaining materials. Based on this, the presence of charcoal is an evidence of the occurrence of current or past fires, contributing with a valuable source of (paleo) environmental proxy data. In this sense, the objective of this work is to assess the physical and chemical changes that occur on scraps of logs from Araucaria columnaris. It will be done during the charring process in muffle, at different temperatures, allowing us to assess important influences in relation to its forming process. The charring temperatures of the logs in natura were in a range of 50 to 1000 ° C, with ranging temperatures from 50 to 50 ° C, and heating ramp of 25 °C min-1. The analysis of this result seeks to allow the proposal of a methodology that assists in the characterization of the natural processes involved in the forming of this type of material, serving as a contribution to (paleo) environmental inferences.

DnaJ proteins or heat shock protein 40s (HSP40s) form one of the largest heat shock protein families. In this study, 2 cDNAs encoding Nile tilapia (Oreochromis niloticus) DnaJ proteins (On-DnaJ B9b and On-DnaJ C3a) were successfully cloned and characterized. The structures and organizations of these two genes are first reported in the present study. On-DnaJ B9b is approximately 2.1 kb long and contains 2 exons and 1 intron, while On-DnaJ C3a is approximately 12 kb long and contains 12 exons and 11 introns. Under normal conditions, On-DnaJ B9b mRNA is highly expressed in gonad and trunk kidney tissues, while On-DnaJ C3a transcripts are abundantly expressed in gills, intestine, liver, and trunk kidney tissues. Following pathogenic infections, the expression of both genes is induced in the liver, spleen and head kidney tissues of Nile tilapia that were infected with two virulent pathogenic bacteria, Streptococcus agalactiae and Flavobacterium columnare. Silencing of these two genes was first carried out, and the results clearly indicated their crucial roles under both heat and bacterial stress conditions. The fundamental knowledge obtained from this study indicates the characteristic basic biofunctions of heat shock proteins in the regulation of intracellular proteins during infection, which involve preventing protein aggregation, promoting protein refolding, and activating unfolded protein degradation.