David J. Pasnik

FIRST REPORT OF STREPTOCOCCUS AGALACTIAE AND LACTOCOCCUS GARVIEAE FROM A WILD BOTTLENOSE DOLPHIN (TURSIOPS TRUNCATUS)

The isolation and characterization of two bacterial species, Streptococcus agalactiae and Lactococcus garvieae, previously unreported in wild marine mammals are described from a freshly dead bottlenose dolphin, Tursiops truncatus, from Kuwait Bay, Kuwait, in September 2001. Conventional and rapid identification systems were used to determine that isolates from muscle and kidney were S. agalactiae and L. garvieae, respectively. The isolates were gram-positive, catalase-negative, oxidase-negative, nonhemolytic cocci. The S. agalactiae was serotyped to group antigen B, whereas the L. garvieae could not be assigned to any serogroup. These Kuwait isolates displayed considerable homogeneity with corresponding American Type Culture Collection (ATCC) type isolates. Although the dolphin S. agalactiae isolate was nonhemolytic, it was biochemically similar to S. agalactiae isolated from mullet sampled in the concurrent Kuwait Bay fish kill. Some biochemical heterogeneity was observed between the dolphin isolates and corresponding mammalian ATCC type isolates, especially with Voges Proskauer, alaninephenylanaline-proline arylamidase, and alpha-galactosidase tests. Nile tilapia, Oreochromis niloticus, experimentally infected with the dolphin S. agalactiae and L. garvieae isolates experienced 90% and 0% mortalities, respectively. This is the first isolation of S. agalactiae and L. garvieae from a wild marine mammal, and the microbial characteristics established here provide pertinent information for the future isolation of these bacteria.

Un-ionized Ammonia Exposure in Nile Tilapia: Toxicity, Stress Response, and Susceptibility to Streptococcus agalactiae

Abstract A series of experiments were conducted to determine the toxicity, behavior, blood glucose stress response, and disease susceptibility in Nile tilapia Oreochromis niloticus following un-ionized ammonia (UIA) exposure. The acute toxicity of un-ionized ammonia to Nile tilapia was measured in a 96-h static test. The median lethal concentration (LC50) was 1.46 mg/L UIA at 24 and 48 h postexposure, 1.33 mg/L at 72 h postexposure, and 0.98 mg/L at 96 h postexposure. No mortalities were noted in unexposed (0 mg/L) control fish or fish exposed to 0.5 mg/L UIA. However, 93–100% mortalities were observed within 24 h among fish exposed to 2.0, 3.0, or 4.0 mg/L UIA. In additional UIA exposure experiments, Nile tilapia were exposed to sublethal concentrations (0.32–0.37 mg/L UIA) for 24 h and then administered an intraperitoneal injection with 750 colony-forming units (CFU) of Streptococcus agalactiae per fish. Mortalities of UIA-exposed and control fish were not significantly different 21 d postchallenge. Blo...

A commercial rapid optical immunoassay detects Streptococcus agalactiae from aquatic cultures and clinical specimens.

The BioStar STREP B Optical ImmunoAssay (STREP B OIA) (BioStar OIA Strep B Assay Kit; BioStar Incorporation, Louisville, CO, USA), commonly used for diagnosis of human maternal group B streptococcus (GBS) colonization, was evaluated for its diagnostic and analytical sensitivity and specificity to aquatic animal GBS isolates, cross-reactivity, and diagnosis and recovery of GBS directly from clinically- infected fish swabs. STREP B OIA identified 25 known fish and dolphin GBS isolates. Thirteen non-GBS negative control isolates from fish and other animals were negative, giving 100% analytical specificity and no cross-reactivity. Three groups of 6 Nile tilapia (Oreochromis niloticus) (mean weight of 40.60+/-1.70 g) each were inoculated intraperitoneally with either 10(6) colony-forming units (cfu) GBS/fish, 10(6) cfu Streptococcus iniae/fish or 100 microL of tryptic soy broth (TSB) and observed for mortality for 7 days. The nare and brain of all fish were swabbed and subjected to the STREP B OIA for detection of GBS antigen immediately after swabbing (0 h) or 24, 48 and 72 h post-swabbing and compared to conventional culture on trypticase soy agar with 5% sheep blood. The STREP B OIA method demonstrated a diagnostic sensitivity of 75.0% and a diagnostic specificity of 69.2% compared to direct TSA. The percent agreement between OIA and culture was 100%. GBS antigen could be retrieved by OIA following 72-h storage of swabs. These results demonstrate the utility of the STREP B OIA to identify GBS from culture and directly from swabs of clinically- infected fish.

Pathogenicity of Streptococcus ictaluri to Channel Catfish

The pathogenicity of a Streptococcus ictaluri isolate in channel catfish Ictalurus punctatus at the fry (0.5 g), fingerling (15 g), and juvenile (55 g) stages was determined by experimental bath immersion and injection experiments. Channel catfish were exposed in 1-L immersion baths containing 10(8), 10(9), 10(10), 10(11) or 10(12) colony-forming units (cfu) of S. ictaluri. Fish were also injected intraperitoneally with 0.1 mL of bacterial solution for final doses of 10(4), 10(5), 10(6), 10(7), or 10(8) cfu of S. ictaluri per fish. Streptococcus ictaluri caused mortality in fry, fingerling, and juvenile channel catfish within 21 d postinfection. When mortalities were calculated based on size and challenge route, the cumulative percent mortalities were 11% for fry and 0% for fingerlings by the bath immersion route and 14% for fingerlings and 6% for juveniles by the injection route. Isolation of S. ictaluri from moribund and dead catfish was confirmed by the newly established BIOLOG profile (MicroLog3 system). The results indicate that channel catfish were only susceptible to high concentrations of S. ictaluri and that juvenile channel catfish were less susceptible, possibly explaining why little mortality has been attributed to S. ictaluri infection in catfish aquaculture.

Specific serum antibody responses in channel catfish (Ictalurus punctatus) provide limited protection against Streptococcus ictaluri challenge.

Passive immunization studies were conducted to determine the role of specific antibodies in immunity to Streptococcus ictaluri. Adult channel catfish (Ictalurus punctatus) were injected i.p. with tryptic soy broth as control or with 1.5 × 10(7)colony-forming units (cfu) S. ictaluri/fish at 0, 30, and 60 d, and serum was collected 90 d after the original challenge. Fish were passively immunized by i.p. injection with serum from the tryptic soy broth (TSB) control group, anti-S. ictaluri serum from fish immunized three times and sampled at 90 d (SSI), or heat-inactivated anti-S. ictaluri serum from fish immunized three times and sampled at 90 d (HISSI). These passively immunized fish were then challenged 72 h later with 1.5 × 10(8)cfu S. ictaluri/fish. Over 21 d, the mean cumulative percent survival was 43.3 (TSB), 63.3 (SSI), and 50.0 (HISSI). A significant difference in cumulative percent survival was noted between the TSB and the HISSI groups, and significant differences were noted between these groups and the SSI group. Serum obtained from immunized fish 72 h after passive immunization exhibited increased anti-S. ictaluri antibody levels. Twenty-one days after the challenge, the HISSI and SSI group antibody levels significantly increased above their corresponding pre-challenge levels. No significant (r(2)=0.0806; P<0.5985) correlation between increased pre-challenge specific serum antibody levels and survival after challenge was demonstrated when analyzing the control and passive immunization groups. The results indicate that both specific anti-S. ictaluri antibodies and non-specific immune responses are important for protection against S. ictaluri.

Bacillus licheniformis isolated during a fish kill is non-pathogenic

= 7,meanweight = 18.2 g)atfishkillsitesinthe river. Dead fish captured and examined in theareaexhibitedanadvancedstateofautolysis,sonodead fish were used in this study.The approximatemean water quality parameters at the samplingsites were: temperature 20.22°C, dissolved oxygen8.26 mg/L, salinity 7.6 and pH 8.36. Blood glucoseat the time of capture was measured using themethods of Evans