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The efficacy of florfenicol for control of mortality associated with Edwardsiella icatluri was studied in fingerlings of Channel Catfish Ictalurus puntatus (Delta strain), Blue Catfish I. furcatus (DB strain), and a hybrid catfish (Delta strain Channel Catfish × DB strain Blue Catfish). On day 0, fish were immersion challenged in 65-L aquaria. For each of the three species of catfish, 10 aquaria were randomly assigned to two treatment groups, either treated with florfenicol at 0 mg/kg of body weight (unmedicated feed) or at 10 mg/kg (medicated feed). Fish were treated for 10 consecutive days, monitored for mortality during this treatment period, and observed for 14 d afterwards. Post observation, all survivors were humanely euthanized in tricaine methanesulfonate, cultured for E. ictaluri, and examined for gross pathology. The mean cumulative percent mortality from enteric septicemia of catfish (ESC) challenge among the three genotypes of catfish did not differ between Blue Catfish, hybrid, and Channel Catfish in treated or control groups. However, the florfenicol-treated fish had a significantly lower mean cumulative mortality (6%) than the controls (78%). All genotypes of catfish tested were responsive to treatment with florfenicol-medicated feed for control of mortality associated with ESC. There were no significant differences in mortality associated with hybrid catfish, blue catfish, and Channel Catfish (Delta strain).

Aquaflor is a feed premix for fish containing the broad spectrum antibacterial agent florfenicol (FFC) incorporated at a ratio of 50% (w/w). To enhance the effectiveness of FFC for salmonids infected with certain isolates of Flavobacterium psychrophilum causing cold water disease, the FFC dose must be increased from the standard 10 mg·kg⁻¹ body weight (BW)·d⁻¹ for 10 consecutive days. A residue depletion study was conducted to determine whether FFC residues remaining in the fillet tissue after treating fish at an increased dose would be safe for human consumption. Groups of Rainbow Trout Oncorhynchus mykiss (total n = 144; weight range, 126-617 g) were treated with FFC at 20 mg·kg⁻¹ BW·d⁻¹ for 10 d in a flow-through system (FTS) and a recirculating aquaculture system (RAS) each with a water temperature of ∼13°C. The two-tank RAS included a nontreated tank containing 77 fish. Fish were taken from each tank (treated tank, n = 16; nontreated tank, n = 8) at 6, 12, 24, 48, 72, 120, 240, 360, and 480 h posttreatment. Florfenicol amine (FFA) concentrations (the FFC marker residue) in skin-on fillets from treated fish were greatest at 12 h posttreatment (11.58 μg/g) in the RAS and were greatest at 6 h posttreatment (11.09 μg/g) in the FTS. The half-lives for FFA in skin-on fillets from the RAS and FTS were 20.3 and 19.7 h, respectively. Assimilation of FFC residues in the fillets of nontreated fish sharing the RAS with FFC-treated fish was minimal. Florfenicol water concentrations peaked in the RAS-treated tank and nontreated tanks at 10 h (453 μg/L) and 11 h (442 μg/L) posttreatment, respectively. Monitoring of nitrite concentrations throughout the study indicated the nitrogen oxidation efficiency of the RAS biofilter was minimally impacted by the FFC treatment.

The efficacy of Aquaflor (florfenicol; FFC) to control mortality caused by Streptococcus iniae in tilapia was evaluated under field conditions. The trial was initiated following presumptive diagnosis of S. iniae infection in a mixed group of fingerling (mean, 4.5 g) Nile Tilapia Oreochromis niloticus and a hybrid of Nile Tilapia×Blue Tilapia O. aureus. Diagnoses included mortality in source tank; examination of clinical signs and presence or absence of gram-positive cocci in brain, and collection of samples for microbiological review and disease confirmation of 60 moribund fish. Following presumptive diagnosis, tilapia (83/tank) were randomly transferred to each of 20 test tanks receiving the same water as the source tank (test tank water was not reused). Tilapia were offered either nonmedicated control feed or FFC-medicated feed (FFC at 15 mg/kg body weight/d; 10 tanks per regimen) for 10 consecutive days followed by a 14-d observation period during which only the nonmedicated control feed was of...

Aquaflor® [50% w w−1 florfenicol (FFC)], is approved for use in freshwater-reared warmwater finfish which include tilapia Oreochromis spp. in the United States to control mortality from Streptococcus iniae. The depletion of florfenicol amine (FFA), the marker residue of FFC, was evaluated after feeding FFC-medicated feed to deliver a nominal 20 mg FFC kg−1 BW d−1 dose (1.33× the label use of 15 mg FFC kg−1 BW d−1) to Nile tilapia O. niloticus and hybrid tilapia O. niloticus × O. aureus held in a recirculating aquaculture system (RAS) at production-scale holding densities. Florfenicol amine concentrations were determined in fillets taken from 10 fish before dosing and from 20 fish at nine time points after dosing (from 1 to 240 h post-dosing). Water samples were assayed for FFC before, during and after the dosing period. Parameters monitored included daily feed consumption and biofilter function (levels of ammonia, nitrite and nitrate). Mean fillet FFA concentration decreased from 13.77 μg g−1 at 1-h post dosing to 0.39 μg g−1 at 240-h post dosing. Water FFC concentration decreased from a maximum of 1400 ng mL−1 at 1 day post-dosing to 847 ng mL−1 at 240 h post-dosing. There were no adverse effects noted on fish, feed consumption or biofilter function associated with FFC-medicated feed administration to tilapia.

Francisella noatunensis subsp. orientalis (Fno) (syn. F. asiatica) is an emergent Gram-negative facultative intracellular bacterium. Although it is considered one of the most pathogenic bacteria in fish, there are no commercially available treatments or vaccines. The objective of this project was to determine the most efficacious concentration of florfenicol (FFC) [10, 15 or 20 mg FFC kg(-1) body weight (bw) per days for 10 days] administered in feed to control experimentally induced infections of Fno in Nile tilapia, Oreochromis niloticus (L.), reared in a recirculating aquaculture system. The cumulative mortality of fish that received 0, 10, 15 or 20 mg FFC kg(-1) bw per day was 60, 37, 14 and 16%, respectively. Francisella noatunensis subsp. orientalis genome equivalents were detected in water from all challenged groups with slight reduction in the concentration in the florfenicol-treated groups 4 days after treatment. The mean LOG of CFU Fno mg(-1) spleen was 3-5 and was present in all challenged groups at necropsy 11 days after treatment (21 days after challenge). Results show that florfenicol administered at doses of 15 and 20 mg FFC kg(-1) bw per days for 10 days significantly reduced mortality associated with francisellosis in Nile tilapia.

The safety of Aquaflor® (50% w/w florfenicol [FFC]) incorporated in feed then administered to tilapia for 20 days (2× the recommended duration) at 0, 15, 45, or 75 mg/kg body weight/day (0, 1, 3, or 5× the recommended dose of 15 mg FFC/kg BW/d) was investigated. Mortality, behavioral change, feed consumption, body size, and gross and microscopic lesions were determined. Estimated delivered doses were >96.9% of target. Three unscheduled mortalities occurred but were considered incidental since FFC-related findings were not identified. Feed consumption was only affected during the last 10 dosing days when the 45 and 75 mg/kg groups consumed only 62.5% and 55.3% of the feed offered, respectively. There were significant, dose-dependent reductions in body size in the FFC-dose groups relative to the controls. Treatment-related histopathological findings included increased severity of lamellar epithelial hyperplasia, increased incidence of lamellar adhesions, decreased incidence of lamellar telangiectasis in the gills, increased glycogen-type and lipid-type hepatocellular vacuolation in the liver, decreased lymphocytes, increased blast cells, and increased individual cell necrosis in the anterior kidney, and tubular epithelial degeneration and mineralization in the posterior kidney. These changes are likely to be of minimal clinical relevance, given the lack of mortality or morbidity observed. This study has shown that FFC, when administered in feed to tilapia at the recommended dose (15 mg FFC/kg BW/day) for 10 days would be well tolerated.

A dose titration study was conducted to determine the dosage of florfenicol (FFC) in feed to control Streptococcus iniae-associated mortality in Nile tilapia Oreochromis niloticus. Six tanks were assigned to each of five treatments: (1) not challenged with S. iniae and fed unmedicated feed; (2) challenged with S. iniae by injection and fed unmedicated feed; (3) challenged with S. iniae and given FFC at 5 mg/kg of body weight (bw) in medicated feed; (4) challenged with S. iniae and given 10 mg FFC/kg bw; and (5) challenged with S. iniae and given 15 mg FFC/kg bw. Treatment was initiated the day after inoculation, and feed was administered for 10 d. Cumulative mortality was 0% in the unchallenged, untreated group; 35.8 +/- 4.4% (mean +/- SE) in the challenged, unmedicated group; 19.2 +/- 2.7% in the 5-mg/kg treated group, 12.5 +/- 3.8% in the 10-mg/kg group, and 2.5 +/- 1.1% in the 15-mg/kg group. The cumulative mortality was significantly less in each challenged, FFC-treated group than in the challenged, unmedicated controls (5 mg/ kg: P = 0.0156; 10 mg/kg: P = 0.0007; 15 mg/kg: P0.0001). The efficacy of the 10- and 15-mg/kg FFC dosages was studied in a separate dose confirmation study. Fish in all tanks were injected with S. iniae. At 4 h postinoculation, 10 tanks were assigned to each of three feed treatments: (1) unmedicated feed; (2) 10 mg FFC/kg bw; and (3) 15 mg FFC/kg bw. Cumulative mortality was 20.5 +/- 2.0% in the challenged, unmedicated group; 11.0 +/- 2.1% in the 10-mg/kg group; and 5.5 +/- 2.4% in the 15-mg/kg group. Mortality was significantly less in the medicated groups than in the challenged, unmedicated control group (10 mg/kg: P = 0.0270; 15 mg/kg: P = 0.0007). Fish in both studies were necropsied, cultured for bacteria, and examined for gross lesions. The minimum inhibitory concentration of FFC against S. iniae in both studies ranged from 0.5 to 1.0 microg/mL. Florfenicol was palatable, safe, and efficacious for control of Nile tilapia mortality due to S. iniae infection.

The studied in channel catfish Ictalurus punctatus fingerlings held in 80-L aquaria. Nonabraded fish were challenged by immersion on day 0. Thirty 80-L tanks were randomly assigned in equal numbers to two treatment groups, one in which fish were fed a commercial diet without florfenicol (unmedicated feed) and one in which they were fed a diet containing 10 mg of florfenicol/kg of body weight (medicated feed) for ten consecutive days. Mortality was monitored during the treatment period and a 14-d posttreatment observation period. At the end of the posttreatment period, all fish were euthanized, examined for gross lesions, and cultured for F. columnare. Significantly fewer fish fed the medicated diet died (8.0%) than fish fed the unmedicated diet (54.2%). Flavobacterium columnare was cultured from 15.0% of the medicated fish, compared with 68.9% of the unmedicated fish. The gross lesions in the fish were consistent with columnaris disease, and F. columnare was cultured from 99.5% of the dead fish. No differences were observed in weight gain and appetence between the medicated and unmedicated groups. For the F. columnare strain used in this study, the minimal inhibitory concentration of florfenicol ranged from 0.5 to 1.0 mg/mL in the 30 bacterial cultures obtained from infected fish, and the mean disk diffusion zone of inhibition was 40 mm. There were no adverse effects among the medicated fish. Administration of florfenicol at a dosage of 10 mg/kg body weight for 10 d was efficacious and safe for the control of mortality from F. columnare infection in channel catfish.

The efficacy of emamectin benzoate (SLICE) against sea lice infestations of Atlantic salmon, Salmo salar L., is typically assessed using untreated fish, or fish treated with alternative therapeutants, as controls. The State of Maine, USA, is currently under active management for the OIE-notifiable pathogen, infectious salmon anaemia virus (ISAV); consequently, neither control group is feasible in this region. Untreated salmon risk extensive damage from the ectoparasites, and threaten to increase vector-borne exposure or susceptibility of farms to ISAV; and the only treatment presently available in Maine is SLICE. However, because sea lice infestations are unlikely to resolve spontaneously, and response to treatment occurs within weeks, use of a pretreatment baseline is a reasonable alternative for confirmatory studies. We evaluated SLICE efficacy on Atlantic salmon farms in Cobscook Bay 2002-2005, in the absence of untreated controls, using pretreatment lice loads as a reference for calculation. Maximum efficacy ranged from 68% to 100% reduction from initial levels. Time-to-maximum efficacy ranged from 1 to 8 weeks after treatment initiation. Efficacy duration, measured between first reduction and first progressive rise in counts, ranged from 4 to 16 weeks.

A field study to assess the efficacy of florfenicol (FFC) against enteric septicemia of catfish (ESC) was conducted with pond-reared channel catfish fingerlings held in 0.1-acre earthen ponds. Fish were challenged with Edwardsiella ictaluri in a natural pond outbreak or by cohabitation with E. ictaluri-infected fish held in netpens. Fourteen ponds were assigned in equal number to two treatment groups, that is, either treated (with 10 mg FFC/kg body weight in medicated feed) or not treated (control) for 10 consecutive d. The threshold for enrollment into the study was 0.3% cumulative mortality attributed to ESC. Treatment was initiated on different dates for each pond because each pond was enrolled when 33 fish/pond were diagnosed with ESC based on clinical signs, lesions, or positive cultures. Mortality was monitored during the 10-d treatment period and during a 14-d posttreatment observation period. At the end of the 14-d posttreatment observation period, all fish were euthanized, and 20 fish from each pond were examined by gross necropsy and evaluated for the presence of E. ictaluri by bacterial culture. The odds of a mortality in the control group were 2.20 times the odds of a mortality in the FFC-treated group. Significantly fewer (P≤ 0.05) FFC-medicated catfish died in comparison to unmedicated catfish. The minimum inhibitory concentration of FFC for this strain of E. ictaluri was 0.25 μg/mL in all fish that were assayed. The mean zone of inhibition (Kirby Bauer) was 36.8 mm from E. ictaluri isolates of test fish. There were no FFC treatment-related lesions seen on gross pathology. FFC was efficacious and safe for control of mortality from E. ictaluri infection in catfish.

Florfenicol is a broad-spectrum antibiotic currently used in salmon aquaculture and proposed for use in channel catfish. A residue depletion study was performed under commercial aquaculture conditions to determine a withdrawal period. Market weight catfish (∼ 2 lb) maintained in a 0.1 acre test pond at a 7000 fish/acre stocking density were dosed with commercially formulated florfenicol in feed for 12 days at an average rate of 9.3 mg/kg b.w./day over the last 10 days, or 93% of the proposed dose. Twenty fish each were collected 1, 2, 4, 7, 14, and 21 days post withdrawal of medicated feed and levels of the marker residue, florfenicol amine (FFA), in muscle tissue were determined via a validated HPLC-UV method. By 4 days average muscle FFA levels were below the proposed 1000 ppb tolerance for FFA and beyond this time point all individual fish were below this level.

SLICE™ (active ingredient 0.2% emamectin benzoate (EMB)), a feed premix developed by Schering-Plough Animal Health for the control of sea lice on cultured salmonids, is registered for use in several countries and is being prescribed on an emergency basis in Canada and the United States. The concentration of EMB in feed administered to farmed salmon ranges from 1 to 25 μg g−1. To determine the acute toxicity of the compound to juvenile and adult American lobster (Homarus americanus), commercial salmon feed was coated with SLICE™ at a range of concentrations and provided to the animals for 7 d in the laboratory. The LC50 is estimated to be 644 μg g−1 (95% CI=428, 1275) for adult lobsters and >589 μg g−1 for stage V and VI juvenile lobsters. The consumption of medicated pellets by adult lobsters decreased significantly with increasing concentration of EMB. Adult lobsters that died during the study had a significantly greater concentration of emamectin B1a in their muscle tissue than those that survived. These results support the conclusion that salmon feed medicated with EMB at the concentrations used by the aquaculture industry is unlikely to pose an acute lethal threat to adult and small juvenile American lobsters.

–A dose titration study was conducted to determine the appropriate dosage of florfenicol in feed to control mortality in channel catfish Ictalurus punctatus associated with enteric septicemia of catfish caused by Edwardsiella ictaluri. Six tanks (20 fish/ tank) were assigned to each of the following treatment: 1) not challenged with E. ictaluri and fed unmedicated feed; 2) challenged with E. ictaluri and fed unmedicated feed; 3) challenged and fed 5-mg florfenicol/kg body weight (kg bw); 4) challenged and fed 10-mg florfenicol/kg bw; or 5) challenged and fed 15-mg florfenicol/kg bw. Treatment was initiated the day after inoculation, and feed was administered by hand at 2.5% body weight for 10 consecutive days. Feeding activity was scored for all groups and was noted to be significantly less than the challenged, unmedicated group. Cumulative mortality in the challenged untreated group was 60%. The mortality in the unchallenged untreated group was 0%, and in die 5-, 10-, 15-mg florfenicol/kg bw group was 2.5%, 0.8%, and 2.5%, respectively. The mortality in each challenged, treated group and the non-challenged control group was significantly less than the challenged, unmedicated controls (P < 0.0001 for each contrast). There were no pairwise statistically significant contrasts among the florfenicol treated groups and the non-challenged control group. All 600 fish in the study were necropsied, cultured for bacteria, and examined by gross pathology. No specific lesions that could be associated with the antibiotic were observed. The efficacy of the 10 mg/kg dosage was confirmed in a separate dose confirmation study. In this study, fish in 30 tanks (20 fish/ tank) were infected with E. icraluri by immersion. Two days post-inoculation, fish in 15 tanks were hand-fed unmedicated feed, and 15 tanks were hand-fed medicated feed at a dosage of 10-mg florfenicol/kg bw at 2.5% body weight for 10 d. Feeding activity was scored and was noted to be significantly less than the challenged, unmedicated group. Cumulative mortality in the florfenicol group (14%) was significantly less than cumulative mortality in the untreated group (87.3%) (P < 0.0001). All 600 fish were submitted for bacterial culture, necropsied. and examined for gross pathology, and once again, no specific lesions that could be associated with the antibiotic were observed. The minimum inhibitory concentration of florfenicol against E. ictaluri in both studies was 0.25 ug/mL. Florfenicol was palatable, safe, and efficacious for control of mortality due to infection by E. ictaluri in catfish.

A tolerance study was conducted to determine the palatability of florfenicol to channel catfish Ictalurus punctatus. Four tanks of fish (20 fish/tank) were assigned to each of five treatments distinguished by the amount of florfenicol given in feed per kilogram of body weight, namely, 0, 10, 20, 40, or 100 mg. Fish were fed at a rate of 2.5% of body weight per day for 10 consecutive days. On day 11, all surviving fish were euthanatized, counted, and weighed as a group. Florfenicol- medicated feed was palatable to fish at doses of 10, 20, 40, and 100 mg for 10 consecutive days. All 400 fish were necropsied and examined by histopathology, and no treatment-related changes were observed. In a separate exploratory efficacy study, four tanks (20 fish/tank) were assigned to each of the following treatments: (1) not challenged with Edwardsiella ictaluri and fed unmedicated feed, (2) challenged with E. ictaluri and fed unmedicated feed, (3) challenged with E. ictaluri and fed florfenicol at 10 mg per kilo...

The safety and efficacy of emamectin benzoate, administered in-feed to Atlantic salmon smolts, Salmo salar L., held in freshwater, was evaluated as a preventative treatment against sea lice, Lepeophtheirus salmonis, following transfer of fish to seawater. In the safety study, salmon smolts held in freshwater were fed with diets containing emamectin benzoate at nominal doses of 0 (control), 50 (recommended dose) and 250 (5× recommended dose) μg kg−1 fish day−1 for 7 days (days 0–6). Actual dose rates, based on measured concentrations of emamectin benzoate in feed, differences in fish weight, and feed consumed, were 0, 54, and 272 μg kg−1 day−1, respectively. On day 9, fish were transferred to seawater and observed for 14 days. No differences in feeding response, coordination, behaviour, gross and histological appearance were observed between control fish and those that received 54 μg kg−1 day−1. Among smolts that received 272 μg kg−1 day−1, approximately 50% exhibited darker coloration, and one fish (1%) exhibited uncoordinated swimming behaviour. No pathognomonic signs of emamectin benzoate toxicity were identified. In the efficacy study, smolts held in freshwater were fed an unmedicated ration (control group) or emamectin benzoate at 50 μg kg−1 day−1 (treated group) for 7 days (days 0–6). On day 9, fish were re-distributed to eight seawater tanks, each holding 30 control and 30 treated fish. On days 28, 56, 77 and 109, respectively, control and treated fish in two tanks were challenged with L. salmonis copepodites. When lice in each group reached chalimus stage IV, fish were sampled and the numbers of lice were recorded. Fish challenged at day 109 were sampled for the second time when lice were at the adult stage. Efficacy was calculated as the reduction in the mean number of lice on treated fish relative to the mean on control fish. Treatment with emamectin benzoate resulted in an efficacy of 85.0–99.8% in fish challenged at days 28–77, from the start of treatment, and lice counts were significantly lower (P

Epidemiological cut-off values were developed for application to antibiotic susceptibility data for Flavobacterium psychrophilum generated by standard CLSI test protocols. The MIC values for ten antibiotic agents against Flavobacterium psychrophilum were determined in two laboratories. For five antibiotics, the data sets were of sufficient quality and quantity to allow the setting of valid epidemiological cut-off values. For these agents, the cutoff values, calculated by the application of the statistically based normalized resistance interpretation method, were

The effect of 0.2% emamectin benzoate (SLICE; Intervet/ Schering-Plough Animal Health, Roseland, New Jersey) administered in top-dressed, pelleted commercial fish feed was evaluated for control of freshwater Argulus sp. in goldfish Carassius auratus and koi carp, a variant of common carp Cyprinus carpio, in freshwater aquaria at 24-25 degrees C. Sixteen individually housed goldfish were each exposed to 37 Argulus. The number of fish lice attached to each fish at the start of the experiment was not determined; however, the total number of motile fish lice in each aquarium (on fish and in the water) was determined at the start and end of each experiment. Eight goldfish were fed the control diet (0 microg x kg fish biomass(-1) x d(-1)) and eight were fed the medicated diet (50 microg x kg fish biomass(-1) x d(-1)) for seven consecutive days. After treatment, fish louse infestation in controls was 20.5 +/- 1.5 (mean +/- SE) lice per fish. No Argulus were found on fish in the treated group. In a separate experiment, 10 individually housed koi were each exposed to 128 Argulus. Five koi were fed the control diet and five were fed a low-dose medicated diet (5 microg x kg fish biomass(-1) x d(-1)) for 7 d. After treatment, fish louse infestation among the controls was 14.6 +/- 3.8 lice per koi. No Argulus were found on koi in the treated group. Hence, a 7-d regimen of oral emamectin benzoate controlled experimental infestation of Argulus when administered to goldfish at 50 microg x kg fish biomass(-1) x d(-1) and to koi at 5 microg x kg fish biomass(-1) x d(-1).

Francisella asiatica is a recently described, Gram-negative, facultative intracellular fish pathogen, known to be the causative agent of francisellosis in warm-water fish. Francisellosis outbreaks have increased in frequency among commercial aquaculture operations and have caused severe economic losses in every case reported. The lack of effective treatments for piscine francisellosis led us to investigate the potential efficacy of florfenicol for inhibition of F. asiatica in vitro and as an oral therapeutic agent in vivo . The MIC of florfenicol for F. asiatica , as determined by the broth dilution method, was 2 μg/ml, which indicates its potential efficacy as a therapeutic agent for treatment of francisellosis. The intracellular susceptibility of the bacterium to florfenicol in tilapia head kidney-derived macrophages (THKDM) was also investigated. Addition of florfenicol to the medium at 10 μg/ml was sufficient to significantly reduce bacterial loads in the THKDM in vitro . Cytotoxicity assays done in infected THKDM also demonstrated drug efficacy in vivo , as determined by lactate dehydrogenase (LDH) release. Levels of LDH released from infected THKDM were significantly lower in macrophages treated with florfenicol ( P < 0.001) than in untreated cells. In medicated-feed trials, fish were fed 15 mg of florfenicol/kg of fish body weight for 10 days, and the feeding was initiated at either 1, 3, or 6 days postchallenge. Immersion challenges resulted in 30% mean percent survival in nontreated fish, and fish receiving medicated feed administered at 1 and 3 days postinfection showed higher mean percent survival (100% and 86.7%, respectively). A significant decrease ( P < 0.001) in bacterial numbers (number of CFU/g of spleen tissue) was observed in treated groups compared to nontreated infected fish at both 1 and 3 days postchallenge. There were no differences in bacterial burden in the spleens between fish treated 6 days postchallenge and untreated controls. In conclusion, if florfenicol is administered during early stages of infection, it has the potential for effectively treating piscine francisellosis, including the capacity for intracellular penetration and bacterial clearance.

In vitro studies were conducted to assess the sensitivity of Edwardsiella ictaluri, the etiological agent of enteric septicemia of catfish (ESC), to the antibacterial drug florfenicol (FFC). Twelve different E. ictaluri isolates from cases submitted between 1994 and 1997 to the Thad Cochran National Warmwater Aqua-culture Center fish diagnostic laboratory (Stoneville, MS) were used for testing. These isolates originated from channel catfish ( Ictalurus punctatus) infected with E. ictaluri through natural outbreaks of ESC in the commercial catfish ponds in Mississippi. Seven hundred sixty-seven additional cultures of E. ictaluri were obtained from channel catfish infected experimentally with E. ictaluri. In some of these experimental infections, FFC was used for treatment. These cultures of E. ictaluri were identified by morphological and biochemical tests. Kirby-Bauer zones of inhibition (in mm) for FFC against E. ictaluri were determined using standard methods. The minimum inhibitory concentration (MIC) of FFC was determined for the natural outbreak E. ictaluri isolates and arbitrarily selected experimental cultures. The zones of inhibition for FFC tested with E. ictaluri ranged from 31 to 51 mm. The MIC for FFC tested with E. ictaluri was consistently 0.25 μg/ml. Edwardsiella ictaluri tested in these studies were highly sensitive to FFC in vitro.

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.

A topically applied 65% permethrin spot-on (Defend EXspot Treatment for Dogs, Schering-Plough Animal Health) used for flea and tick control on dogs was evaluated for repellency and efficacy against the yellow fever mosquito, Aedes aegypti, a vector of canine filariasis. Six dogs were randomly assigned to receive a single application of 65% permethrin on Day 0 (n=3) or to remain untreated as controls (n=3). Dogs were anesthetized and exposed to 100 unfed, female mosquitoes in screened cages for 2 hours on Days -6, -4, -1, 0, 1, 7, 14, 21, and 28. Mosquito landing rates, engorgement rates, and mortality were determined for each mosquito challenge. Cages were thoroughly cleaned after each mosquito challenge. Treatment of dogs with 65% permethrin reduced the mosquito landing rates by 96.3% 6 hours after treatment and by 82.5% on Day 1. Mosquito mortality, relative to the control group, was 28.2% 6 hours after treatment, ranged from 84.0% to 90.9% through Day 21, and declined to 50.3% 28 days after treatment. Successful feeding by mosquitoes was significantly (P=.05) reduced on Days 1 through 28. The 65% permethrin spot-on treatment killed and repelled significantly (P =.05) more mosquitoes on treated dogs versus untreated dogs for 28 days after treatment.

The efficacy of two formulations of a topically applied 65% permethrin spot-on (Defend Exspot Treatment for Dogs, Schering-Plough Animal Health) was evaluated against experimental infestations of the cat flea Ctenocephalides felis and the lone star tick Amblyomma americanum in dogs. Eighteen dogs were randomly assigned to treatment with 65% permethrin in either diethylene glycol monomethyl ether (DGME; original formulation) or propylene glycol monomethyl ether (PGME) or to be untreated as a control. Treated dogs received either 1 (body weight15 kg) or 2 ml (body weightor =15 kg) of the assigned formulation on Day 0. One hundred unfed, adult C. felis were placed on each dog on Days -6, -1, 4, 11, 18, 25, and 32. Fifty unfed, adult ticks were placed on each dog on Days -1, 3, 9, 16, 23, and 30. Live fleas and ticks were counted and removed on Days 3, 7, 14, 21, and 28. Treatment of dogs with the 65% permethrin in DGME reduced flea numbers by 90.4% to 99.9% from Days 3 through 21 (Por =.05) and by 48.2% 28 days after treatment. Treatment of dogs with 65% permethrin in PGME reduced flea numbers by 93.7% to 99.7% from Days 3 through 28 and by 78.4% 35 days after treatment (Por =.05). Treatment with 65% permethrin in DGME reduced tick numbers by 90% or more only on Day 7, whereas treatment with 65% permethrin in PGME reduced the number of live ticks by 90%or more on Days 7 and 14 and approached 90%(87.9%) on Day 21. Efficacy against fleas and ticks for the PGME formulation was significantly better (Por =.05) than for the DGME formulation on Day 28. Findings in this study indicate that both the DGME and PGME formulations of 65% permethrin performed well in reducing numbers of live C. felis and A. americanum on laboratory beagles; however, the PGME formulation was effective approximately 1 to 2 weeks longer than the DGME formulation.

Twenty-five dogs were evenly and randomly allocated by weight to five treatment groups: untreated control, 2 ml 65% permethrin (dogs weighing 15 to 29 kg, "average dogs"), and 2, 3, or 4 ml 65% permethrin (dogs weighingor = 30 kg, "big dogs"). Each dog was infested with 125 unfed, adult cat fleas, Ctenocephalides felis, and 50 unfed, adult brown dog ticks, Rhipicephalus sanguineus, on Days -3 (ticks only), -2 (fleas only), 3, 7, 14, 17, 21, 24, and 28. Fleas and ticks were counted 1 and 3 or 4 days after each infestation. The duration of efficacy (defined as90%) against C. felis was 28 to 31 days. The efficacy against fleas 31 days after application of 2, 3, or 4 ml on big dogs ranged from 79.1% (2 ml) to 100% (4 ml). Big dogs that received either 3 or 4 ml of 65% permethrin had significantly (Por = .05) fewer fleas at several evaluations between 15 and 31 days after treatment. The duration of efficacy against R. sanguineus was 15 (2 ml for big dogs) to 28 (2 ml for average dogs) days. The efficacy against R. sanguineus 28 days after treatment ranged from 79.1% (2 ml on big dogs) to 94.1% (2 ml on average dogs). Significantly (Por = .05) fewer ticks were present at several evaluations after treatment on big dogs that received 3 or 4 ml than were present on big dogs treated with 2 ml. No significant differences were detected between the 3- and 4-ml groups from Days 10 to 31; however, the geometric mean number of ticks in the group treated with 4 ml was numerically lower than that for dogs treated with 3 ml on several occasions. These data indicate that a dose volume of 3 or 4 ml of 65% permethrin is needed to obtain an adequate level and duration of efficacy against both C. felis and R. sanguineus on dogs weighing 30 kg or more.

Four studies were conducted to determine the efficacy of a 1% lambdacyhalothrin pour-on (Saber Pour-on, Schering-Plough Animal Health) for control of sucking lice (Linognathus vituli, Haematopinus eurysternus, Solenopotes capillatus) and biting lice (Damalinia bovis) on beef cattle. Seventy-four mixed-breed cattle naturally infested with one or more species of lice at locations in Wisconsin, Nebraska, Oklahoma, and North Dakota were included in the study. Pretreatment lice samples were taken and identified by genus with the exception that sucking lice were not identified by genus at the North Dakota site. In January or February, half of the cattle at each location received a single application of 1% lambdacyhalothrin pour-on at 10 ml per head for cattle weighing less than 273 kg (600 lb) or 15 ml per head for cattle weighing 273 kg or more. The other cattle at each site served as untreated controls. Lice on designated body areas were counted 2, 4, 6, and 8 weeks after treatment, and the sum of all lice observed on each counting area was reported for each animal. The collective efficacy 6 weeks after treatment against three species of sucking lice (L. vituli, H. eurysternus, S. capillatus) was 88.4% at the Wisconsin site, 92.0% at the North Dakota site, and 100% at the Nebraska and Oklahoma sites. The 1% lambdacyhalothrin pour-on eliminated all biting lice within 2 weeks after treatment, and no biting lice were detected 8 weeks after treatment. A single treatment of 1% lambdacyhalothrin pour-on administered when lice populations were highest (January or February) provided effective season-long control of both biting and sucking lice on cattle.

The efficacy of two formulations of a topically applied 65% permethrin spot-on for dogs (Defend EXspot Treatment for Dogs, Schering-Plough Animal Health Corp.) was evaluated against experimental infestations of the cat flea, Ctenocephalides felis, and the brown dog tick, Rhipicephalus sanguineus. Thirty dogs were randomly allocated to treatment with 65 % permethrin in diethylene glycol monomethyl ether (original formulation), 65 % permethrin in propylene glycol monomethyl ether (test formulation), or to an untreated control group. Dogs assigned to treatment with a permethrin formulation received either 1 or 2 ml of the formulation in accordance with label directions on Day 0. One hundred unfed, adult cat fleas and 50 unfed, adult ticks were placed on each dog on Days -1, 5, 12, 19, 26, 33, and 40. Live fleas and ticks were counted on each dog on Days 2, 7, 14, 21, 28, 35, and 42. Treatment of dogs with either formulation of 65 % permethrin significantly (P.05) reduced the number of live fleas and ticks from Days 2 through 42. No statistical differences were noted between the formulations regarding efficacy against C. felis or R. sanguineus.

Two topically applied spot-on products used for flea and tick control on dogs, 65% permethrin (Defend EXspot Treatment for Dogs, Schering-Plough Animal Health Corp., Union, NJ) and selamectin (Revolution [United States] or Stronghold [Europe], Pfizer Animal Health, New York, NY), were evaluated for repellency and efficacy against Ixodes ricinus, the primary vector of Lyme disease in Europe. Eighteen dogs were evenly and randomly allocated to the following treatments: 1) 65% permethrin, 2) selamectin, 3) untreated control. Dogs were treated by topical application of the assigned product in accordance with product label directions on Day 0. At 7, 14, 21, 28, and 35 days after treatment, each dog was exposed for 2 hours to 50 unfed, adult ticks in a cage with a carpet that covered approximately 70% of the floor area. After the exposure period, dogs were removed from the cages and live and dead ticks were counted on the dogs and in the cages. The number of live ticks recovered was reduced by 90.3% to 99.5% for dogs treated with 65% permethrin (P.0001 versus controls and selamectin), compared with 10.9% to 31.1% for dogs treated with selamectin (P.05 versus controls). The repellency of 65% permethrin was 63.4% to 80.2% against I. ricinus ticks (P.0001 versus controls, P.0007 versus selamectin), compared with 0% to 10.9% repellency for selamectin (P.05 versus controls).

1. Pilocarpine-induced saliva of the soft tick Ornithodoros moubata inhibits platelet aggregation induced by ADP or collagen, even when diluted 2000 times into platelet rich plasma. 2. Saliva contains apyrase (ATP-diphosphohydrolase) activity, which has an optimal pH of 7.0 for ADP and of 8.0 for ATP hydrolysis, respectively. Both Ca2+ and Mg2+ activate the reactions. 3. The mean specific activities for ATP and ADP hydrolysis at pH 7.5 were 0.97 and 0.74 mumoles orthophosphate/min/mg protein. 4. These results, which demonstrate for the first time such activities in the saliva of soft ticks, support the hypothesis that the saliva of blood sucking arthropods serves an anti-hemostatic role during feeding and that large amounts of salivary apyrase activity have evolved independently in hematophagous arthropods.

The objective of this study was to develop a highly specific enzyme-linked immunosorbent assay (ELISA) for the serological detection of anti-Ornithodoros tick antibodies in animals. Affinity-purified rabbit anti-Ornithodoros IgG antibodies were employed in indirect competitive inhibition ELISA assays designed to measure the anti-Ornithodoros antibody titers in other animal species using the domestic goat (Capra hircus) as a large animal model. Repeated infestation of goats with Ornithodoros coriaceus was found to elicit the formation of antibodies capable of inhibiting the binding of the Ornithodoros-specific rabbit IgG. Western blot analysis of goat and rabbit anti-tick antisera demonstrated both animal species to respond immunologically to a set of 9 major protein bands in O. coriaceus salivary gland extracts. The results of these experiments demonstrate that a history of animal exposure to O. coriaceus may be detected serologically by competitive inhibition ELISA.

Three laboratory colonies of the argasid tick Onithodoros moubata porcinus van der Merwe were started from collections made in 1983 at three different sites in Zimbabwe. All of the colonies contained ticks infected with African swine fever (ASF) virus that was readily transmitted by bite to domestic pigs. Although they were maintained on virus-free pig blood, ASF virus infections persisted in the colonies for at least 1 yr. Despite the fact that ASF virus passes transstadially, sexually, and transovarially in this tick species sometime during the following year, the virus disappeared from the colonies. Studies comparing fecundity in infected and uninfected lots of O. moubata porcinus showed that mortality rates were considerably higher among the infected ticks. A similar study with Ornithodoros erraticus Lucas, a tick that harbors and transmits ASF virus on the Iberian Peninsula, gave essentially the same results. This is probably a factor involved in the clearance of ASF virus from tick populations that are not subjected to reinfection. How this information may be applied in the eradication of African swine fever in Portugal and Spain is discussed.