Mark P. Gaikowski

Fish gut microbiota analysis differentiates physiology and behavior of invasive Asian carp and indigenous American fish

Gut microbiota of invasive Asian silver carp (SVCP) and indigenous planktivorous gizzard shad (GZSD) in Mississippi river basin were compared using 16S rRNA gene pyrosequencing. Analysis of more than 440 000 quality-filtered sequences obtained from the foregut and hindgut of GZSD and SVCP revealed high microbial diversity in these samples. GZSD hindgut (GZSD_H) samples (n=23) with >7000 operational taxonomy units (OTUs) exhibited the highest alpha-diversity indices followed by SVCP foregut (n=15), GZSD foregut (n=9) and SVCP hindgut (SVCP_H) (n=24). UniFrac distance-based non-metric multidimensional scaling (NMDS) analysis showed that the microbiota of GZSD_H and SVCP_H were clearly separated into two clusters: samples in the GZSD cluster were observed to vary by sampling location and samples in the SVCP cluster by sampling date. NMDS further revealed distinct microbial community between foregut to hindgut for individual GZSD and SVCP. Cyanobacteria, Proteobacteria, Actinobacteria and Bacteroidetes were detected as the predominant phyla regardless of fish or gut type. The high abundance of Cyanobacteria observed was possibly supported by their role as the fish’s major food source. Furthermore, unique and shared OTUs and OTUs in each gut type were identified, three OTUs from the order Bacteroidales, the genus Bacillariophyta and the genus Clostridium were found significantly more abundant in GZSD_H (14.9–22.8%) than in SVCP_H (0.13–4.1%) samples. These differences were presumably caused by the differences in the type of food sources including bacteria ingested, the gut morphology and digestion, and the physiological behavior between GZSD and SVCP.

Persistence of DNA in carcasses, slime and avian feces may affect interpretation of environmental DNA data

The prevention of non-indigenous aquatic invasive species spreading into new areas is a goal of many resource managers. New techniques have been developed to survey for species that are difficult to capture with conventional gears that involve the detection of their DNA in water samples (eDNA). This technique is currently used to track the invasion of bigheaded carps (silver carp and bighead carp; Hypophthalmichthys molitrix and H. nobilis) in the Chicago Area Waterway System and Upper Mississippi River. In both systems DNA has been detected from silver carp without the capture of a live fish, which has led to some uncertainty about the source of the DNA. The potential contribution to eDNA by vectors and fomites has not been explored. Because barges move from areas with a high abundance of bigheaded carps to areas monitored for the potential presence of silver carp, we used juvenile silver carp to simulate the barge transport of dead bigheaded carp carcasses, slime residue, and predator feces to determine the potential of these sources to supply DNA to uninhabited waters where it could be detected and misinterpreted as indicative of the presence of live bigheaded carp. Our results indicate that all three vectors are feasible sources of detectable eDNA for at least one month after their deposition. This suggests that current monitoring programs must consider alternative vectors of DNA in the environment and consider alternative strategies to minimize the detection of DNA not directly released from live bigheaded carps.

Safety of Aquaflor (Florfenicol, 50% Type A Medicated Article), Administered in Feed to Channel Catfish, Ictalurus punctatus

Aquaflor, a feed premix containing the broad spectrum antibacterial agent florfenicol (50% w/w), is being developed for use to control enteric septicemia (ESC) in channel catfish Ictalurus punctatus caused by the gram-negative enterobacterium Edwardsiella ictaluri. The recommended dose of Aquaflor to control ESC is 10 mg/kg body weight (BW)/day for 10 days. The study objective was to determine the safety of Aquaflor administered in feed to channel catfish at doses of 0 (control), 10, 30, and 50 mg/kg BW/day for 20 consecutive days. Parameters evaluated included daily mortality, behavioral (appetite, distribution, flight/fright response), and water chemistry observations, initial and terminal weight measurements, and gross and microscopic pathology. Medicated feed consumption was 67—86% of target with group mean doses of 8.5 mg/kg BW/day, 24.6 mg/kg BW/day, and 34.9 mg/kg BW/day. There were no mortalities or clinically observable changes noted at any of the dose levels tested. Aquaflor-related changes were limited to the food consumption and histopathology data. Although Aquaflor-related decreased feed consumption was noted in the 30 and 50 mg/kg BW/day groups, there were no differences in fish growth among the treatment groups. Aquaflor-related histopathology findings were limited to a histomorphologically evident dose-dependent decrease in hematopoietic/lymphopoietic tissue in the anterior kidneys, posterior kidneys, and spleens of channel catfish.

Importance of Analytically Verifying Chemical Treatments

Abstract Hydrogen peroxide is considered a low regμLatory priority compound by the U.S. Food and Drug Administration. It is used to control fungal infections on fish eggs. We studied the treatment profiles of hydrogen peroxide in Heath, McDonald egg jar, and Clark–Williamson incubators during treatments intended to deliver an effective regimen of at least 500 μL hydrogen peroxide/L (i.e., treatments of 500 and 1,000 μL/L) for 15 min. Hydrogen peroxide concentrations decreased with increasing distance from the influent water in both Heath and Clark–Williamson incubators. The top treatment tray (tray 2) of the Heath incubator received more than 90% of the intended regimen during the 500 μL/L treatment, whereas at 1,000 μL/L, all trays had hydrogen peroxide concentrations at or above 500 μL/L for 15 min. None of the compartments in the Clark–Williamson incubator received the intended therapeutic regimen when treated at 500 μL/L. The McDonald egg jar system distributed the intended concentration for the desig...

Toxicity of Hydrogen Peroxide Treatments to Rainbow Trout Eggs

Abstract Hydrogen peroxide treatments of 0, 500, 1,000, and 3,000 μL/L, concentrations that were multiples of the Low Regulatory Priority limit of 500 μL/L, were administered for 15 min every weekday (Monday–Friday) to eggs of rainbow trout Oncorhynchus mykiss and steelhead (anadromous rainbow trout) to determine the margin of safety existing for standard egg treatments. All untreated and treated eggs remained free of fungal infection throughout incubation. Hydrogen peroxide treatment reduced the mean percent hatch of rainbow trout eggs by 1.4–5.9% among those treated at 500 μL/L, 6.8–15.4% among those treated at 1,000 μL/L, and 13.2–25.3% among those treated at 3,000 μL/L. Mean percent hatch of rainbow trout eggs treated at 1,000 μL H2O2/L was 7% lower than that for eggs treated at 500 μL H2O2/L. Mean percent hatch of Skamania strain steelhead was significantly reduced by hydrogen peroxide treatment, whereas the mean percent hatch of Ganaraska strain steelhead was similar to the mean percent hatch of rai...

Chronic toxicity of hydrogen peroxide to Daphnia magna in a continuous exposure, flow-through test system

A flow-through, continuous exposure test system was developed to expose Daphnia magna to an unstable compound. 35% Perox-Aid is a specially formulated hydrogen peroxide (a highly oxidative chemical) product approved for use in U.S. aquaculture and therefore has the potential to be released from aquaculture facilities and pose a risk to aquatic invertebrates. The study objective was to assess the effects of 35% Perox-Aid on an aquatic invertebrate by evaluating the survival, growth, production, and gender ratio of progeny from a representative aquatic invertebrate continuously exposed to 35% Perox-Aid. The study design consisted of 6 treatment groups (10 test chambers each) with target hydrogen peroxide concentrations of 0.0, 0.32, 0.63, 1.25, 2.5, and 5.0 mg L(-1). The study was initiated with <24-h-old Daphnia (1 daphnid per chamber) that were exposed to hydrogen peroxide for 21 days. Hydrogen peroxide concentrations < or =1.25 mg L(-1) had no significant effect on Daphnia time to death compared to controls and no significant effect on the time to first brood production and the number of broods produced. Concentrations < or =0.63 mg L(-1) had no significant effect on the total number of young produced. Concentrations > or =0.32 mg L(-1) had a negative effect on Daphnia growth. Hydrogen peroxide had no significant effect on the gender ratio of young produced. All second generation Daphnia were female. A continuous discharge of hydrogen peroxide into aquatic ecosystems is not likely to affect cladocerans if the concentration is maintained at < or =0.63 mg L(-1) for less than 21 days.

Carbon Dioxide as a Tool to Deter the Movement of Invasive Bigheaded Carps

AbstractNonnative bigheaded carps are established in the Mississippi River and there is substantial concern about their potential entry into the interconnected Laurentian Great Lakes. While electrical barriers currently exist as a preventative measure, there is need for additional control mechanisms to promote barrier security through redundancy. We tested the effectiveness of infused carbon dioxide gas (CO2) as a tool to influence the movement and behavior invasive bigheaded carps, namely Bighead Carp Hypophthalmichthys nobilis and Silver Carp H. molitrix, as well as native Bigmouth Buffalo Ictiobus cyprinellus, Channel Catfish Ictalurus punctatus, Paddlefish Polyodon spathula, and Yellow Perch Perca flavescens in an experimental pond. Individuals were monitored with acoustic telemetry before, during, and after CO2 addition to the pond. We noted distinct changes in fish behavior following CO2 addition. Each species except Paddlefish maintained farther distances from the CO2 infusion manifold relative to ...

Use of hydrogen peroxide during incubation of landlocked fall chinook salmon eggs in vertical-flow incubators.

Abstract Six different hydrogen peroxide treatment regimes were evaluated in a series of three trials with landlocked fall Chinook salmon Oncorhynchus tshawytscha eggs incubated in vertical-flow incubators. Six daily 15-min hydrogen peroxide treatment regimes (1,000 mg/L; 1,000 mg/L with a decrease to 500 mg/L during estimated blastopore formation; 2,000 mg/L; 2,000 mg/L with a decrease to 500 mg/L during estimated blastopore formation; 2,500 mg/L; and 2,500 mg/L with a decrease to 500 mg/L during estimated blastopore formation) were compared with daily 15-min treatments of 1,667 mg/L of formalin. Mortality at egg eye-up and fry hatch and from eye-up to hatch was significantly greater in eggs receiving the 2,500-mg/L hydrogen peroxide treatments throughout incubation and in those receiving 2,500 mg/L hydrogen peroxide with a decrease to 500 mg/L during blastopore formation than in either of the 1,000-mg/L hydrogen peroxide treatment regimes or the formalin-treated eggs in the first trial. No significant d...

Effects of Immobilization by Electricity and MS-222 on Brown Trout Broodstock and Their Progeny

Abstract To determine the effects of electrically and chemically induced immobilization on postspawn broodstock and their progeny, age-2 and age-3 female broodstock and age-2 male broodstock of brown trout Salmo trutta were immobilized with electricity or tricaine methanesulfonate (MS-222), stripped of their eggs or milt, and weighed. Eggs taken from electrically immobilized females were fertilized with milt taken from age-2 males that were immobilized with electricity, and eggs taken from females immobilized with MS-222 were fertilized with milt taken from age-2 males that were immobilized with MS-222. After spawning, the mortality and weight of broodstock were compared twice over a 6-month period. Egg viability and growth of offspring fry from each treatment group were also compared. Electricity induced complete and consistent immobilization in brown trout broodstock. Electrically immobilized fish were more easily handled than fish immobilized with MS-222; however, electrically immobilized fish survival...

Efficacy of Hydrogen Peroxide versus Formalin Treatments to Control Mortality Associated with Saprolegniasis on Lake Trout Eggs

Abstract We compared the efficacy of hydrogen peroxide versus formalin treatments to control fungal infections on eggs of lake trout Salvelinus namaycush incubated at a hatchery in Wisconsin. Four strains of lake trout eggs were incubated in six vertical-flow Heath incubators; three replicate incubators for each chemical. Each incubator had 13 trays containing approximately 25,000–30,000 eggs/tray. Formalin (1,667 mg/L) or hydrogen peroxide (1,000 mg/L) treatments were administered once daily for 15 min up to the development of visible eye spots in the eggs (eyed egg stage). Eyed and dead eggs were separated using a photoelectric egg sorter, and the number of live and dead eggs was determined volumetrically. In the hydrogen peroxide test group, the bottom trays of each incubator had fungus present on the eggs, whereas no fungus was observed on eggs treated with formalin. The mean percentage of eyed eggs for an incubator treated with formalin (75%) was significantly greater than an incubator treated with h...