Investigating the Ability of Edwardsiella ictaluri and Flavobacterium covae to Persist within Commercial Catfish Pond Sediments under Laboratory Conditions.

Two prevalent bacterial diseases in catfish aquaculture are enteric septicemia of catfish and columnaris disease caused by Edwardsiella ictaluri and Flavobacterium covae, respectively. Chronic and recurring outbreaks of these bacterial pathogens result in significant economic losses for producers annually. Determining if these pathogens can persist within sediments of commercial ponds is paramount. Experimental persistence trials (PT) were conducted to evaluate the persistence of E. ictaluri and F. covae in pond sediments. Twelve test chambers containing 120 g of sterilized sediment from four commercial catfish ponds were inoculated with either E. ictaluri (S97-773) or F. covae (ALG-00-530) and filled with 8 L of disinfected water. At 1, 2, 4-, 6-, 8-, and 15-days post-inoculation, 1 g of sediment was removed, and colony-forming units (CFU) were enumerated on selective media using 6 × 6 drop plate methods. E. ictaluri population peaked on Day 3 at 6.4 ± 0.5 log10 CFU g-1. Correlation analysis revealed no correlation between the sediment physicochemical parameters and E. ictaluri log10 CFU g-1. However, no viable F. covae colonies were recovered after two PT attempts. Future studies to improve understanding of E. ictaluri pathogenesis and persistence, and potential F. covae persistence in pond bottom sediments are needed.

The Use of Kaolin as a Prophylactic Treatment to Prevent Columnaris Disease (Flavobacterium covae) in Commercial Baitfish and Sportfish Species.

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 effects of novel alginate-lauric arginate coatings with temperature on bacterial quality, oxidative stability, and organoleptic characteristics of frozen stored chicken drumsticks.

Although alginate has been reported to be used as an edible film and coating in food, to the best of our knowledge, this study is the first to investigate the individual effects of coatings, temperatures, storage times, as well as their interactions, on the bacterial quality, deterioration criteria, and sensory attributes of chicken drumsticks. To accomplish this, six groups of chicken drumsticks were treated with different coatings, temperatures, and storage conditions. The groups included 2 % alginate at 25 °C (Alg25) and 50 °C (Alg50), a mixture of 2 % alginate and 2 % LAE at 25 °C (M25) and 50 °C (M50), besides control untreated samples in distilled water at 25 °C (C25) and 50 °C (C50). The chicken drumsticks were stored at -18 °C for 3 months. The results showed that all treated chicken drumsticks induced a significant reduction in all bacterial counts, as well as a significant decrease in pH and thiobarbituric acid values, with an improvement in all sensory attributes, particularly in the M25 and M50 coated samples compared to the C25 and C50. Furthermore, exposing alginate and LAE to a temperature of 50 °C can increase their antimicrobial activity. In conclusion, the innovative combinations of LAE and alginate can be used successfully to decontaminate chicken carcasses in poultry processing plants.

Persistence of a Wild-Type Virulent Aeromonas hydrophila Isolate in Pond Sediments from Commercial Catfish Ponds: A Laboratory Study.

Virulent Aeromonas hydrophila (vAh) is a major bacterial pathogen in the U.S. catfish industry and is responsible for large-scale losses within commercial ponds. Administering antibiotic feeds can effectively treat vAh infections, but it is imperative to discern new approaches and better understand the mechanics of infection for this bacterium. As such, the persistence of vAh in pond sediments was determined by conducting laboratory trials using sediment from four commercial catfish ponds. Twelve chambers contained sterilized sediment, vAh isolate ML-09-119, and 8 L of water maintained at 28 °C and were aerated daily. At 1, 2, 4, 6, and 8 days, and every 7th day post-inoculation for 28 days, 1 g of sediment was removed, and vAh colony forming units (CFU) were enumerated on ampicillin dextrin agar. Viable vAh colonies were present in all sediments at all sampling periods. The vAh growth curve peaked (1.33 ± 0.26 × 109 CFU g-1) at 96 h post-inoculation. The population plateaued between days 14 and 28. No correlations were found between CFU g-1 and physiochemical sediment variables. This study validated the ability of vAh to persist within pond sediments in a laboratory setting. Further research on environmental factors influencing vAh survivability and population dynamics in ponds is needed.

Thermal performance of the electron transport system Complex III in seven Alabama fishes.

Management of fish populations for conservation in thermally variable systems requires an understanding of the fish's underlying physiology and responses to thermal stress. Physiological research at the organismal level provides information on the overall effects of stressors such as extreme temperature fluctuations. While experiments with whole organisms provide information as to the overall effects of temperature fluctuations, biochemical assays of thermal stress provide direct results of exposure that are both sensitive and specific. Electron transport system (ETS; Complex III) assays quantify a rate-limiting step of respiratory enzymes. Parameters that can be estimated via this approach include optimum thermal temperature (Topt ) and optimal breadth of thermal performance (Tbreadth ), which can both be related to organismal-level temperature thresholds. We exposed enzymes of seven fish species (native fish chosen to represent a typical community in Alabama streams) to temperatures in the range 11-44°C. The resultant enzymatic thermal performance curves showed that Topt , the lower temperature for enzyme optimal thermal performance (Tlow ), the upper temperature for enzyme optimal thermal performance (Tup ), and Tbreadth differed among species. Relationships between enzymatic activity and temperature for all fish followed a pattern of steadily increasing enzyme activity to Topt before gradually decreasing with increasing temperature. A comparison of our enzyme optimum and upper-temperature limit results versus published critical thermal maxima values supports that ETS Complex III assays may be useful for assessing organismal-level thermal tolerance.

Evaluating Stress-Mediated Microbial Pathogenesis in Golden Shiners, Notemigonus crysoleucas.

Flavobacterium covae (columnaris) is a microbial pathogen of the Golden Shiner (Notemigonus crysoleucas), a principal bait species. We investigated the effects of density and water temperature on the survival of fish subjected to a columnaris challenge and whether flow cytometry (FCM) could be a fast and reliable method to distinguish and enumerate F. covae populations from water and fish in experimental tanks. Juvenile Golden Shiners averaging 2.62 (±0.78 S.D.) g (negative for F. covae) were used in simultaneous trials at 22°C and 28°C in two ultra-low flow-through systems: each consisting of four treatments and five replicates per treatment. Treatments were fish stocked at either 600 fish/m3 or 2,400 fish/m3 and either challenged with F. covae or not; survival was observed for 48 h after challenge. Samples of water and fish tissue were obtained for FCM enumerations and validation by qPCR. No significant differences in survival were recorded between density treatments; however, high temperature and columnaris challenge treatments showed significantly higher mortality. Bacterial enumeration (number/mL) by FCM highly correlated with bacterial counts r = 0.81 (p = 0.001) in the water samples. Higher water temperatures may have increased columnaris infections and mortality in Golden Shiners. Flow cytometry is a reliable method of enumerating F. covae from experimental tank water samples.

The synergistic effects of plant polysaccharide and Pediococcus acidilactici as a synbiotic additive on growth, antioxidant status, immune response, and resistance of Nile tilapia (Oreochromis niloticus) against Aeromonas hydrophila.

This study evaluated the growth performance, immune responses, and disease resistance of Nile tilapia upon pistachio hulls derived polysaccharide (PHDP) and Pediococcus acidilactici (PA) separately or as synbiotic. Fish received four types of diets: T1, control; T2, PHDP (0.1%); T3, PA (0.2%); T4, PHDP (0.1%) +PA (0.2%) for 56 days. The results showed that final weight and weight gain were markedly higher in fish fed T4 diet than that given T1 and T2 diets (P ≤ 0.05). In addition, a significantly greater specific growth rate was obtained by the T4 diet compared to the control. Fish survival was significantly improved in all supplemented diets compared to the control. On the other hand, the activities of lipase, protease, and amylase showed significant increases in the T4 group compared with other feeding groups. The total leucocytes and lymphocytes proportion significantly elevated in T3 and T4 than remaining groups (P ≤ 0.05). Further, fish fed T3 diet presented significantly higher serum total protein, total immunoglobulin, lysozyme activity (LYZ), alternative complement activity (ACH50), and alkaline phosphatase activity compared to fish fed T1 and T2 diets, while the mentioned indices were found significantly highest in T4 group than others. Fish received T3 and T4 diets had higher skin mucus LYZ and ACH50 than those fed T1 and T2 diets (P ≤ 0.05). The malondialdehyde levels were significantly declined in T3 and T4 when compared to the control. Fish fed T3 and T4 diets demonstrated significantly enhanced superoxide dismutase, catalase, and glutathione peroxidase activities compared to the control. The intestinal propionic acid significantly increased by T2 and T4 diets, while the highest levels of acetic acid detected in fish given T4 diet. The expression levels of tumor necrosis factor α (TNF-α), interleukin 1ß (IL-1ß), and interleukin 10 (IL-10) were significantly affected by T3 and T4 supplements. The efficacy of T4 diet against Aeromonas hydrophila infection was documented by a significantly lower mortality rate. In conclusion, the combination of PHDP and PA presented promising results as a synbiotic feed additive for Nile tilapia.

The implication of autoantibodies in early diagnosis and monitoring of plasmonic photothermal therapy in the treatment of feline mammary carcinoma.

Feline mammary carcinoma (FMC) shows great similarities to human breast cancer in the cellular and molecular levels. So, in cats as in humans, the role of immune responses is indicated to detect and follow up the development of tumors. As a new breast cancer therapeutic approach, Plasmonic Photothermal Therapy (PPTT) is an effective localized treatment for canine and feline mammary-carcinoma. Its systemic effect has not been inquired yet and needs many studies to hypothesis how the PPTT eradicates tumor cells. In this study, it is the first time to detect (P53, PCNA, MUC-1, and C-MYC) feline autoantibodies (AAbs), study the relationship between PCNA AAbs and mammary-tumors, and investigate the effect of PPTT on the humoral immune response of cats with mammary-carcinoma through detection of AAbs level before, during, and after the treatment. The four-AAbs panel was evaluated in serum of normal and clinically diagnosed cats with mammary tumors using Enzyme-Linked Immunosorbent Assay. The panel showed 100% specificity and 93.7% sensitivity to mammary tumors. The panel was evaluated in PPTT monotherapy, mastectomy monotherapy, and combination therapy. PPTT monotherapy decreased AAbs level significantly while mastectomy monotherapy and combination therapy had a nonsignificant effect on AAbs level.

Linking multiple aspects of thermal performance to explore the potential for thermal resource partitioning between a native and an invasive crayfish.

Ecologists require standardized, ecologically relevant information on the thermal ecology of aquatic ectotherms to address growing concerns related to changing climates, altered habitats, and introduced species. We measured multiple thermal endpoints to investigate potential for establishment of the invasive Ringed Crayfish (Faxonius neglectus) in thermally heterogeneous habitat of the narrowly distributed endemic Coldwater Crayfish (Faxonius eupunctus). For each species, we examined the relationships between thermal endpoints at the cellular and organismal levels. We then compared results between the two species to gain insight as to the generality of linkages between cellular and organismal-level endpoints, as well as the potential for thermal niche separation between the native and potential invader. At the cellular level, we found no differences in the temperature for maximum activity of electron transport system enzymes (ETSmax) between species. At the organismal level, F. neglectus preferred significantly warmer temperatures than F. eupunctus, but this difference was small (1.3 °C) and likely to have only limited biological significance. The critical thermal maximum (CTM) did not differ between species. For both species, the thermal performance curve for ETS enzyme activity served as a useful framework to link thermal endpoints and estimate the transition from optimal to stressful temperatures - organismal thermal preference and optimal temperature estimates consistently fell below ETSmax whereas CTM estimates fell above ETSmax. Taken together, the strong similarities in thermal endpoint patterns between the two species suggest habitats thermally suitable for the native F. eupunctus will also be thermally available to expanding populations of F. neglectus, thereby increasing the opportunity for negative interactions and population effects if F. neglectus invades one of the few remaining, uninvaded, critical habitats of F. eupunctus.

Embryonic thermal manipulation of Japanese quail: effects on embryonic development, hatchability, and post-hatch performance.

Embryonic thermal manipulation led to several modifications in molecular, physiological, and biochemical parameters which affect pre- and post-hatch growth performance. The current study aims to elucidate the onset and long-term effects of intermittent thermal manipulations (TM) during two-time windows, early/late, of embryogenesis in Japanese quail (Coturnix japonica) on embryonic development, hatchability, muscle histogenesis, and post-hatch growth performance. Four groups were created; quail eggs in the control group were incubated at 37.7 °C and relative humidity (RH) 55%. Three thermally treated groups were incubated intermittently at 41 °C and 65% RH intermittently (3 h/day): early embryogenesis group (EE) was thermally treated during embryonic days (ED) 6-8, late embryogenesis group (LE) was thermally treated during (ED12-ED14), and early and late embryogenesis group (EL) was thermally manipulated in both time windows. Relative embryo weights in EL and EE were significantly lighter than those in LE and Ctrl groups. The hatched chicks were reared under optimal managemental conditions (three replicates per treatment). Average daily feed intake was recorded, and feed conversion ratio (FCR) was calculated. Histological and quantitative analyses of muscle fibers were performed. The results revealed that TM led to significant hypertrophy of quail breast muscle in (EE). Intermittent short-term (3-6 h) thermal manipulation (39-40 °C) protocols during early embryogenesis (ED6-ED8) could be recommended to enhance muscle mass growth and breast muscle yield in the Japanese quail.

An Epidemiological Study on Orodental Disorders in 3,791 Working Donkeys in Egypt: Prevalence and Risk Factors.

Orodental disorders are common in equidae and can lead to serious clinical complications. This study determines the prevalence rate (PR) of orodental disorders in working donkeys in Egypt and their potential risk factors. During 4 years, 3,791 donkeys were examined in six Egyptian governorates. Full case history and thorough clinical and oral examinations were performed. Radiography and oroendoscopy were undertaken whenever possible. All data were statistically analyzed using Poisson generalized linear models to compare PR among governorates, sex, years, body condition score, and age groups and to determine the potential risk factors. Of 3,791 examined donkeys, 954 donkeys (25.17%) had orodental disorders. The PRs of acquired disorders were 9.81% sharp enamel points, 5.86% buccal ulcers, 4.88% hook, 4.70% overgrown teeth, 4.19% periodontal disease, 3.11% ramp, 2.60% dental caries, 2.37% dental calculus, 2.30% diastema, 2.06% step mouth, 2.00% soft tissue injuries other than buccal ulcers, 1.77% worn tooth, 1.37% loose tooth, 1.29% exaggerated transverse ridge, 1.24% fractured teeth, 0.82% missing teeth, 0.69% wave mouth, 0.50% molar table angle change, and 0.45% wolf tooth overgrowth. The PRs of the congenital disorders were 1.90% displaced teeth, 1.37% deviated teeth, 0.58% retained teeth, 0.16% overbite, 0.16% underbite, 0.11% supernumerary teeth, and 0.11% premolar cap. The risk factors significantly associated (P < .05) with the orodental disorders were age and sex of the animal, geographic location, and year of examination. This study provides a database for future studies on orodental disorders and for designing effective preventive and therapeutic strategies for these disorders in donkeys.

Effects of dietary Lactobacillus plantarum (KC426951) in biofloc and stagnant-renewal culture systems on growth performance, mucosal parameters, and serum innate responses of Nile tilapia Oreochromis niloticus.

Effects of dietary Lactobacillus plantarum (KC426951) on growth and innate responses of Nile tilapia Oreochromis niloticus were evaluated in biofloc technology system and stagnant-renewal culture system (SRCS). The 90-day-long experiment contained four treatments: SRCS without probiotic (T1), SRCS with probiotic (T2), biofloc without probiotic (T3), and biofloc with probiotic (T4). The administration dose of probiotic was 2 × 108 CFU kg-1 diet. At the end of experiment, the mean final weights, specific growth rates, feed conversion ratios, and total biomass were significantly (P < 0.05) better in BFT treatments, with no significant effect of probiotic on these parameters in both culture systems. Meanwhile, skin mucosal parameters including total protein (TP), lysozyme (LYZ), alkaline phosphatase (ALP), and protease (PRO) activity were significantly enhanced following probiotic supplementation. T4 treatment displayed a significantly higher LYZ and ALP activity in mucus versus other treatments. Also, serum alternative complement activity was significantly heightened in probiotic-supplemented fish. Superoxide dismutase activity in T4 was detected higher than that of SRCS groups. The results of the current study demonstrated the enhancement of some mucosal and serum innate responses of Nile tilapia in both culture systems upon L. plantarum (KC426951) supplementation.