Effects of metomidate hydrochloride sedation on blood glucose and marketability of transported threespot gourami Trichogaster trichopterus

Our objectives were to determine whether sedation with metomidate hydrochloride (hereafter, "metomidate") during transportation of threespot gourami Trichogaster trichopterus would prevent an increase in blood glucose levels and improve fish marketability (i.e., based on appearance and behavior) in comparison with unsedated controls. Threespot gourami are obligate air-breathers that possess a labyrinth organ, enabling the fish to respire air above the water surface; these fish should be lightly sedated during transport. Fish were transported for approximately 24 h via truck and domestic airline. Blood was sampled at 0, 2, 6, and 12 h posttransport ation, and appearance and behavior were observed at 0, 2, 4, 6, and 12 h and 7 d posttransportation. Metomidate concentrations tested were 0.0 (control), 0.1, 0.2, 0.3, and 0.4 mg/L. At the concentrations tested, metomidate neither inhibited elevations in blood glucose nor improved marketability. Fish that were transported with 0.3-mg/L metomidate were less marketable based on behavioral indices, and fish that were transported with 0.4-mg/L metomidate had higher glucose levels than control fish. Use of metomidate as a transport sedative for threespot gourami should be considered with caution and may be problematic at the concentrations tested; however, further research examining additional indices of stress may clarify metomidate use for this species.     

Analysis of Short-Term Cortisol Stress Response in Channel Catfish by Anesthetization with Metomidate Hydrochloride and Tricaine Methanesulfonate

The ability of the anesthetics metomidate hydrochloride and tricaine methanesulfonate (MS-222) to mitigate the cortisol stress response of Channel Catfish Ictalurus punctatus was evaluated during a 10-min confinement stress. The cortisol concentrations of Channel Catfish anesthetized in metomidate hydrochloride remained consistent throughout the 10-min exposure; however, for fish anesthetized with MS-222 and nonanesthetized fish, cortisol concentrations were approximately 7- and 22-fold higher, respectively, than the baseline concentrations. While both anesthetics reduced cortisol concentrations relative to those of nonanesthetized fish, these results suggest that MS-222 is an appropriate anesthetic to use during the initial 5 min of sedation and that metomidate hydrochloride is appropriate for longer periods of sedation.

Received November 18, 2014; accepted April 9, 2015     

Effects of Metomidate Hydrochloride Sedation on Blood Glucose and Marketability of Transported Threespot Gourami Trichogaster trichopterus

Abstract

Our objectives were to determine whether sedation with metomidate hydrochloride (hereafter, “metomidate”) during transportation of threespot gourami Trichogaster trichopterus would prevent an increase in blood glucose levels and improve fish marketability (i.e., based on appearance and behavior) in comparison with unsedated controls. Threespot gourami are obligate air-breathers that possess a labyrinth organ, enabling the fish to respire air above the water surface; these fish should be lightly sedated during transport. Fish were transported for approximately 24 h via truck and domestic airline. Blood was sampled at 0, 2, 6, and 12 h posttransportation, and appearance and behavior were observed at 0, 2, 4, 6, and 12 h and 7 d posttransportation. Metomidate concentrations tested were 0.0 (control), 0.1, 0.2, 0.3, and 0.4 mg/L. At the concentrations tested, metomidate neither inhibited elevations in blood glucose nor improved marketability. Fish that were transported with 0.3-mg/L metomidate were less marketable based on behavioral indices, and fish that were transported with 0.4-mg/L metomidate had higher glucose levels than control fish. Use of metomidate as a transport sedative for threespot gourami should be considered with caution and may be problematic at the concentrations tested; however, further research examining additional indices of stress may clarify metomidate use for this species.

Received February 16, 2011; accepted December 18, 2011.     

Characterization of four Flavobacterium columnare (Flexibacter columnaris) strains isolated from tropical fish

Four Flavobacterium columnare strains (AJS 1–4) were isolated from black mollies (Poecilia sphenops) and platies (Xiphophorus maculatus), showing white spots on the back, head and skin ulcers. The isolates developed characteristic rhizoid yellow pigmented colonies on Shieh agar and typical growth in Shieh broth. They were Gram-negative, filamentous bacteria exhibiting flexing movements. When compared to F. columnare strains isolated from temperate fish, it was noted that the four strains originating from tropical aquarium fish are more capable of growing at higher temperatures, the opposite being true for the strains isolated from temperate fish. Biochemical characterization and agglutination tests proved that the isolated strains could be classified as F. columnare. Low minimal inhibitory concentration (MIC) values were found for chloramphenicol, erythromycin, furazolidone, kanamycin, lincomycin, nalidixic acid, oxytetracycline and streptomycin. MIC values were high for colistin, sulfamethoxazole and neomycin. Pathogenicity studies were performed on black mollies. When these animals were submersed in an infective solution of the F. columnare strains, a marked difference in virulence was noted among the four isolated strains, strain AJS 1 being the most virulent one and strain AJS 4 being of low virulence.     

Remote intramuscular injection of immobilising drugs into fish using a laser-aimed underwater dart gun

Sixty coldwater and warmwater fish ranging in weight from 2 to 35 kg were injected intramuscularly with the hypnotics alphaxalone-alphadolone and metomidate hydrochloride and the non-depolarising muscle relaxant gallamine triethiodide using a laser-aimed underwater dart gun. Alphaxalone-alphadolone produced sufficient sedation for easy netting within five to 20 minutes at doses between 0.3 and 0.5 ml/kg, with induction being somewhat faster in warmwater species. The pattern of induction was similar with metomidate but required doses of 40 to 60 mg/kg. The muscle relaxant gallamine triethiodide showed promise as a practical agent for the capture and handling of large fish by virtue of its smooth induction of paralysis at doses between 1 and 3 mg/kg and its reversible supplementation with orally administered metomidate hydrochloride.     

Communications: Tolerance of Rainbow Trout to Dissolved Oxygen Supplementation and a Yersinia ruckeri Challenge

Abstract

Dissolved oxygen accounted for a significant proportion of the variation in mortality rate of rainbow trout Oncorhynchus mykiss acclimated for 10 weeks to a range of dissolved oxygen levels and challenged with Yersinia ruckeri. Moderate levels of oxygen supersaturation (150%) resulted in greater cumulative mortality (17.9%) among fish exposed to a 0.5-h static bath challenge with Y. ruckeri (1 × 10⁷ colony-forming units/mL). In contrast, hypoxic (70%) and normoxic (100%) levels of dissolved oxygen resulted in 12.8% and 10% cumulative mortality. Higher oxygen levels provided no additional “protection” against infection by Y. ruckeri in this study.     

A COMPARISON OF ALFAXALONE AND TRICAINE METHANESULPHONATE (MS-222) IN TWO FISH SPECIES

The purpose of this study was to evaluate the effectiveness of the alfaxalone formulation Alfaxan? as an immersion anesthetic in tropical fish species compared to that of tricaine methanesulfonate (MS-222). 22 black spot barbs (Puntius filamentosis) measuring (mean±SD) 11.4 ±1.4 cm in body length and 22 peacock cichlids (Aulonocara spp.) (measuring 8.4 ± 1.6cm were anesthetized in water baths containing 100 mg/L of MS-222 buffered with 200 mg/L of bicarbonate or 5 mg/L of alfaxalone following a 2-week washout period. Time to maximum effect, recovery periods, self-righting, spontaneous swimming movements, opercular movements, and response to noxious stimuli were recorded. The following results are for the black spot barbs following MS-222 and alfaxalone anesthesia, respectively: mean times (±SD) to surgical anesthesia were 5.5 ± 2.11 and 3.27 ± 1.72 minutes and mean recovery times were 2.95 ± 0.9 and 9.14 ± 3.15 minutes. The peacock cichlid anesthetic protocols for MS-222 (20 of 22 cichlids) and alfaxalone (20 of 21 cichlids) produced the following results, respectively: mean times (±SD) to surgical anesthesia were 14.75 ±5.43 and 11.1 ± 9.84 minutes and mean recovery times were 3.6 ±0.82 and 22.4 ±11.3 minutes. Median recovery time from 5 mg/L alfaxalone was significantly longer (P < 0.001) in both species, by 5 minutes for black spot barbs and by 17 minutes for peacock cichlids. Variation in induction and recovery times between species was observed, with black spot barbs having significantly (P < 0.0001) faster induction times when treated with both drugs, and a faster recovery time from 5 mg/L alfaxalone.     

Effect of Droncit (Praziquantel) on Yellow Grubs Clinostomum marginatum and Eye Flukes Diplostomum spathaceum in Channel Catfish

Abstract

Channel catfish Ictalurus punctatus with infections of yellow grubs Clinostomum marginatum and eye flukes Diplostomum spathaceum were treated with 2 mg Droncit (praziquantel)/L in a water bath for 2 and 4 h. At the time of treatment, 80% of the fish had visible yellow grubs and 70% had opaque eyes. Fourteen days after treatment, 40.1% of the yellow grubs in the 2-h treatment were dead. No grubs were dead in the 4-h treatment; however, the number of yellow grubs was reduced in the treated fish. No dead eye flukes were seen in untreated fish after 14 d, but 59.2 and 44.4% of eye flukes in the 2- and 4-h treatments, respectively, were dead. At 21 d, the percentages of dead eye flukes were 86% (2 h) and 90.1% (4 h). Droncit had limited effect on yellow grubs in channel catfish, but the chemical had a beneficial effect of reducing numbers of eye flukes.     

In vitro ruminal fermentation characteristics of gum arabic under concentrate and forage substrate conditions

Gum arabic (GA) has potential rumen modifier functions. This is the first study to investigate the in vitro ruminal fermentation characteristics of GA. Rumen fluid was collected from ruminal fistulated wethers; rolled barley and ryegrass straw were used as substrates for concentrate and forage conditions, respectively. Besides incubating with the substrates alone (control), GA or potato starch (PS) was added at 0.2%, 1.0%, and 2.0% along with substrates. Under the concentrate substrate condition, GA treatments showed higher total gas production in 24-h incubation, but lower methane production in 24- and 48-h incubation compared with PS treatments (p < 0.05). The 1.0% and 2.0% GA and 0.2% and 1.0% PS treatments showed higher dry matter and neutral detergent fiber digestibility and lower NH₃-N, and higher short chain fatty acid concentrations compared with the control at 24-h incubation (p < 0.05). The GA treatments also showed a lower acetate/propionate ratio than PS treatments at 48-h incubation (p < 0.01). Under the forage substrate condition, the treatment effects were not significant, except for a higher proportion of propionate with GA than with PS at 24 and 48 h of incubations. We thus concluded that GA supplement may exert potential rumen modifier effects particularly under concentrate feeding condition.     

Aspects of animal welfare and species protection in the international trade of ornamental fish and air transport to Germany

The number of ornamental fish kept in German aquariums is rising, but the supply of offspring is not sufficient to meet the needs of the fish enthusiasts. Therefore millions of ornamental fish from foreign countries are being imported to Germany. This provokes a number of new problems regarding the protection of species and the animals' welfare during transportation. For the assessment of the transport conditions, 1000 shipments of ornamental fishes were evaluated at the Rhein-Main-Airport Frankfurt, Germany. Water samples were taken from 100 shipments and were examined for anaesthetics. The results are disturbing: 41% of the shipments have total transportation times between 31-42 h resulting in an oxygen deficit in the transport bags (5.2%) and dead ornamental fish. Also damage of the transportation containers, high fish densities as well as drastic size differences amoung the fish in one transport bag were noticeable. In 99% of the cases, formal defects could be noted. The German association of pet stores has declared that fish belonging to three special fresh water families should not be kept in an aquarium, but 1 200 of such fish were imported. The HPLC analysis of the water samples revealed an additive. The non-declared anaesthetic 2-phenoxyethanol was present in all samples taken from shipments based out of Singapore. The results emphasize that improvements are urgently necessary in the control of the air transport and trade with Ornamental fish.     

Communications: Protective Immunity Induced by a Commercial Vibrio Vaccine in Hybrid Striped Bass

Abstract

Commercial Vibrio anguiliarum-V. ordalii bacterin was used to vaccinate hybrid striped bass (Morone saxatilis ♀ × M. chrysops ♂) to test the vaccine efficacy against vibriosis. Vaccination by direct immersion of fish in diluted Vibrio vaccine for 20 s resulted in increased protective immunity. The relative percent survival of hybrid striped bass challenged 35 d after vaccination was 66.7% for those challenged by 1-h immersion exposure to 7.03 × 10⁷ V. anguillarum cells/mL, 75.0% for those challenged by injection with 3.51 × 10⁵ cells/fish, and 86.7% for those challenged by injection with 3.51 × 10⁴ cells/fish.     

Current status of aquatic veterinary services for ornamental fish in Australasia

ABSTRACT

The ornamental fish industry is growing quickly globally. Asia represents 51% of the export market for this industry and in Singapore alone, it has an estimated market value of US$66 million. Despite the economic importance of this industry for Australasia, aquatic veterinary support for the ornamental fish sector is scarce. In many cases, ornamental fish owners do not seek advice from veterinarians. While the reasons behind this are difficult to elucidate, the low perceived value of fish compared to the cost of consulting a veterinarian, and limited knowledge about the availability of veterinary services for aquatic animals may contribute to the lack of interaction with veterinarians. Considering the growing importance of the ornamental fish industry, more education and data are needed about the services offered by veterinarians with aquatic animal health expertise. Knowledge on aquatic veterinary medicine is limited, reinforcing the need for more educational programmes and dissemination of aquatic veterinary services within the Australasian region.     

Phenotypic and genotypic characteristics of Mycobacterium isolates from fighting fish Betta spp. in Malaysia

Mycobacteriosis due to mycobacteria is one of the most common bacterial diseases in ornamental fish. We describe here the phenotypic and genotypic characteristics of Mycobacterium isolates from fighting fish Betta spp. using ATCC Mycobacterium marinum, Mycobacterium fortuitum and Mycobacterium chelonae as references. A total of four isolates (M1, M2, M3, M4) were obtained from four out of 106 fish samples using selective agar, and identified to Mycobacterium genus using acid-fast staining and 16s rRNA gene-based genus specific polymerase chain reaction. DNA sequencing and NCBI-BLAST analysis further identified isolate M1 as M. marinum and isolates M2, M3, M4 as M. fortuitum. Morphological, physiological and biochemical tests were carried out for phenotypic characterizations. Universal M13 and wild-type phage M13 RAPD dendogram was generated to illustrate the genetic relationship of the isolates and reference strains.     

Effectiveness of Praziquantel Bath Treatments against Bothriocephalus acheilognathi in Grass Carp

Abstract

Many states ban the importation of fish infected with the Asian tapeworm Bothriocephalus acheilognathi. Treatment with an effective tapeworm parasiticide prior to shipment would allow fish to be imported that might otherwise be rejected. In this study, extended praziquantel bath treatments of infected grass carp Ctenopharyngodon idella were tested to determine their efficacy against Asian tapeworms. Preliminary praziquantel treatments of 0.7 mg/L or more for an exposure period of 24 h at grass carp densities of 69 g/L were effective in eliminating all tapeworms from 22-g fish. When the treatment duration was lowered to 12 h, a praziquantel concentration of 2.8 mg/L was required for effective treatment. In replicated studies, significantly reduced tapeworm numbers (0–5 tapeworms per treated fish compared with 30+ tapeworms per control fish) were observed following 24-h exposures to praziquantel at 0.5, 0.7, 1.0, and 1.5 mg/ L. Tapeworm elimination occurred only at the maximum concentration of 1.5 mg/L when 88-g grass carp were stocked at a density of approximately 6 g/L. When fish density was raised to 60 g/L, the 1.5-mg/L treatment did not eliminate all tapeworms: 4 of 30 fish contained a single tapeworm.     

Yolk‐sac larval development of the substrate‐brooding cichlid Archocentrus nigrofasciatus in relation to temperature

In order to conserve and culture the cichlid fish Archocentrus nigrofasciatus, more information about its reproductive biology and its larval behavior and morphogenesis is necessary. Currently, temperatures ranging from 21 to 27 °C are used in ornamental aquaculture hatcheries. Lower temperatures are preferred to reduce the costs of water heating, and 23 °C is usually the selected temperature. However, there is limited information on culturing protocols for ornamental species and most of the information generated on this topic remains scarce. Thus, the present study examines the morphological development of Archocentrus nigrofasciatus during the yolk‐sac period up to the age of 100 h post‐hatching in relation to 2 temperature regimes used in ornamental aquaculture: a temperature of 27 °C (thermal optimum) and a decreased temperature of 23 °C (thermal tolerance). The results of this study suggest that the 27 °C temperature generates intense morphological changes in yolk‐sac development in a shorter period. This has advantages as it reduces the time of yolk‐sac larval development, and, thus, minimizes the transition phase to exogenous feeding and maximizes the efficiency at which yolk is converted into body tissues. The present paper provides necessary information to produce freshwater ornamental fish with better practices so as to increase larval survival and capitalize on time for growth.     

Communications: Isolation of Several Mycobacterium Species from Fish

Abstract

Several species of marine fish caught in the wild and of freshwater ornamental fish were used in this study. Infected organs (liver, spleen, and kidney) were sampled for mycobacteria. Decontaminated tissue samples were plated onto selective media for mycobacterial recovery. After initial isolation, fluorescent and acid-fast staining techniques identified bacterial colonies to genus. Profiles of biochemical growth characteristics were used to further identify the isolates to species. Five species of Mycobacterium were identified: M. simiae, M. scrofulaceum, M. marinum, M. chelonae, and M. fortuitum. Of these, M. simiae and M. scrofulaceum have not been previously reported from fish. Tissue samples containing focal granulomatous lesions were prepared for electron microscopic examination.     

Susceptibility of Juvenile and Yearling Bull Trout to Myxobolus cerebralis and Effects of Sustained Parasite Challenges

Abstract

Evaluation of the susceptibility of bull trout Salvelinus confluentus fry to Myxobolus cerebralis infection using two different laboratory challenge models demonstrated that the method of administering the exposure dose affected infection prevalence. Administration of a low parasite dose (500 per fish) in a single exposure did not establish infection, but when the same cumulative dose was administered over 21 d the prevalence of infection was 45%. The results of challenges at a high exposure dose (5,000 per fish) were similar, infections being detected in 24% of the fish receiving a single dose and 40% of those administered the same dose in multiple exposures. Clinical disease was not detected in fish exposed via either challenge method. The susceptibility of yearling bull trout was tested by means of a single high dose of 10,000 per fish, and infection was detected in only 5% of those fish.     

The efficacy of amoxicillin sodium against streptococcosis in cultured olive flounder Paralichthys olivaceus and its pharmacokinetics

The efficacy of amoxicillin sodium for controlling field and experimental Streptococcus iniae and S. parauberis infections in olive flounder (Paralichthys olivaceus) was evaluated after a single intramuscular administration. Furthermore, the minimal inhibitory concentrations (MIC) against 21 Streptococcus strains were determined. In addition, the pharmacokinetics and residue depletion in olive flounder were investigated. Single intramuscular doses of amoxicillin sodium at 20, 40, 80, and 160 mg/kg b.w. fish significantly reduced cumulative mortality rates to 18.8–31.3% (P < 0.05) for S. iniae and to 5.0–15.0% (P < 0.01) for S. parauberis, whereas the S. iniae‐ and S. parauberis‐infected positive control groups showed cumulative mortality rates of 68.8% and 60.0%, respectively. In a S. parauberis outbreak, amoxicillin sodium reduced the cumulative mortality rate to 7.5% and 4.8% at 20 and 40 mg/kg b.w. fish, respectively, whereas that of the untreated control group was 35.2%. Peak plasma concentrations (C_max) following a single intramuscular dose of 40 and 80 mg/kg b.w. fish were 62.64 (T_max, 1.59 h) and 87.61 (T_max, 3.02 h) μg/mL, respectively, with large AUC_0?t/MIC and C_max/MIC ratios, and sufficient T > MIC (time for maintaining plasma drug concentration greater than MICs) for S. iniae and S. parauberis. The estimated withdrawal period of amoxicillin sodium from muscle of olive flounder was about 8 days at 40 mg/kg b.w. fish (at 22 ± 1 °C). These results demonstrated a single intramuscular administration of amoxicillin sodium to be effective against streptococcosis in olive flounder.     

The association of Flavobacterium columnare strains of high and low virulence with gill tissue of black mollies (Poecilia sphenops).

The ability of a high virulence strain (AJS 1) and a low virulence strain (AJS 4) of Flavobacterium columnare (Flexibacter columnaris) to attach to the gills of black mollies (Poecilia sphenops) was investigated. For that purpose, two groups of 25 black mollies each were immersed in a bath containing 10(6) CFU/ml of F. columnare AJS 1 or AJS 4. At regular intervals from 1 to 12 h after the contact infection, fish were sacrificed and gills, skin, spleen and heart were sampled for bacteriology. Samples of the gills were taken for immunohistochemical and electron microscopic examination. Bacteriological examination proved that the number of gill-associated F. columnare was higher for AJS 1 than for AJS 4. Strain AJS 1 was isolated from the heart and spleen of 6 and 1 of the 16 examined animals, respectively. Strain AJS 4 was not isolated from the internal organs of any fish. When examined immunohistochemically, strain AJS 1 was found closely associated with gill epithelium whereas this was not the case for strain AJS 4. The adherence of bacteria to the gill tissue challenged with the virulent strain AJS 1 was also clearly demonstrated using scanning and transmission electron microscopy. These results indicate that adhesion of F. columnare to the gill tissue constitutes an important step in pathogenesis.     

Physiological responses of horses to 24 hours of transportation using a commercial van during summer conditions

Fifteen mature horses (mares, n = 6); geldings, n = 9) were used to assess the physiological responses of 24 h of transport in a commercial van under California summer conditions. The study was conducted on four consecutive days, and data were collected on d 1 and d 2 to obtain baseline values and to determine any diurnal variation in the individual measurements. Travel commenced on d 3 at 0800 for 24 h, with a total of 1,622 km traveled. Blood samples were collected at 0800, 1100, 1400, 2000, and 0200 each day. Horses were weighed and rectal temperatures recorded at 0800 each day and at 2000 each day except d 3. Body weight, rectal temperature, serum cortisol, serum lactate, and white blood cell (WBC) counts exhibited diurnal variation (P = .0001) on d 1 and d 2. Body weight immediately after unloading showed a 6% loss. At 24 h following transit, a 3% deficiency in body weight loss remained. The WBC counts showed a progressive increase with duration of travel and peaked at the termination of transport. Dehydration measures of hematocrit and total protein increased during transport and returned to baseline during the posttransport period. Serum concentrations of lactate, creatine kinase and aspartate aminotransferase increased during transport and in the early posttransit period, but returned to baseline values at the conclusion of the 24-h posttransport period. Glucose concentration increased with the initiation of transport and did not decrease to baseline concentration at the end of the 24-h posttransport period. Plasma cortisol and neutrophil:lymphocyte ratio increased with duration of transit and returned to baseline during the posttransport period. These data clearly showed physiological responses of horses undergoing 24 h of transport including changes in muscle metabolism, stress indices, dehydration and immune parameters, and body weight. These responses may increase disease susceptibility and influence energy availability for athletic performance following long-term transport of horses.