Jeffrey M. Hinshaw

Multiple antimicrobial resistance in plague: An emerging public health risk

Antimicrobial resistance in Yersinia pestis is rare, yet constitutes a significant international public health and biodefense threat. In 1995, the first multidrug resistant (MDR) isolate of Y. pestis (strain IP275) was identified, and was shown to contain a self-transmissible plasmid (pIP1202) that conferred resistance to many of the antimicrobials recommended for plague treatment and prophylaxis. Comparative analysis of the DNA sequence of Y. pestis plasmid pIP1202 revealed a near identical IncA/C plasmid backbone that is shared by MDR plasmids isolated from Salmonella enterica serotype Newport SL254 and the fish pathogen Yersinia ruckeri YR71. The high degree of sequence identity and gene synteny between the plasmid backbones suggests recent acquisition of these plasmids from a common ancestor. In addition, the Y. pestis pIP1202-like plasmid backbone was detected in numerous MDR enterobacterial pathogens isolated from retail meat samples collected between 2002 and 2005 in the United States. Plasmid-positive strains were isolated from beef, chicken, turkey and pork, and were found in samples from the following states: California, Colorado, Connecticut, Georgia, Maryland, Minnesota, New Mexico, New York and Oregon. Our studies reveal that this common plasmid backbone is broadly disseminated among MDR zoonotic pathogens associated with agriculture. This reservoir of mobile resistance determinants has the potential to disseminate to Y. pestis and other human and zoonotic bacterial pathogens and therefore represents a significant public health concern.

GnRHa-induced ovulation of brown trout (Salmo trutta) and its effects on egg quality

Abstract The effectiveness of a GnRH analogue (GnRHa) for inducing ovulation in brown trout was examined, and the effects of the maternal hormone-treatment on egg quality and larval development were evaluated. Two injections of 10 μg/kg body weight of GnRHa given 3 days apart effectively induced ovulation. Within 6 days from the second injection, 80% of the injected fish had ovulated compared to only 10% of the control fish. Mean time to ovulation was reduced significantly (P

Effects of Illumination and Prey Contrast on Survival and Growth of Larval Yellow Perch Perca flavescens

Abstract The influence of illumination and prey contrast on survival, growth, and feeding of larval yellow perch fed Artemia sp. nauplii was tested in an experimental upflow culture system. The treatment with high light level and high prey contrast (205 lux, black background) supported the best survival (mean, 44.2%) and growth (mean total length, 8.9 mm; mean wet weight, 5.0 mg) from hatching to age 14 d. Mean survival was 37.2% when light was low and prey contrast high (75 lux, black background). In the remainder of the tests, survival to age 14 d ranged from 0 to 6.9%. Low light and low prey contrast caused a 1–2 d delay in initial feeding. Visibility of the prey resulting from contrast with the surroundings, rather than light level, accounted for most of the variation in the ability of yellow perch larvae to acquire adequate food.

Thyroid hormones in brown trout (Salmo trutta) reproduction and early development

Gravid brown trout (Salmo trutta) females were injected with various doses of a synthetic gonadotropin-releasing hormone analog (GnRHa), given with or without an injection of triiodothyronine (T3), in order to investigate the potential of T3 (a) to enhance the stimulatory effect of GnRHa on ovulation, and (b) to enhance the growth and survival of the produced progeny. From the time the hormonal treatments were initiated until ovulation was detected 5–38 days later, endogenous plasma T3 levels increased from an average of 3.6 to 11.6 ng ml−1. Injection with 20 mg T3 kg−1 body weight, further elevated plasma T3 levels at ovulation (16.0 ng ml−1. Mean time to ovulation was reduced significantly in fish injected with 10 μg kg−1 of GnRHa, whereas treatment with lower doses was ineffective. Injection with T3 did not enhance the ovulatory response of brown trout to GnRHa. Unfertilized eggs obtained from T3-injected females had a higher T3 content, suggesting a transfer of T3 from the maternal circulation into the oocytes. Maternal T3 injection had no effect on egg fertilization rates, embryo survival to eyeing and hatching, or the prevalence of abnormal larvae at the time of hatching. Length and weight gain of the progeny during yolk absorption was also not influenced by maternal T3 treatment. At the completion of yolk-sac absorption, progeny from females injected with T3 had a higher prevalence of skeletal abnormalities than controls. The results suggest that in teleosts like brown trout, which have high endogenous circulating T3 levels, treatment of females with T3 does not enhance responsiveness to GnRHa and it has the potential for deleterious effects on their offspring.

Independent Emergence of Yersinia ruckeri Biotype 2 in the United States and Europe

ABSTRACT Biotype 2 (BT2) variants of the bacterium Yersinia ruckeri are an increasing disease problem in U.S. and European aquaculture and have been characterized as serovar 1 isolates that lack both peritrichous flagella and secreted phospholipase activity. The emergence of this biotype has been associated with an increased frequency of enteric redmouth disease (ERM) outbreaks in previously vaccinated salmonid fish. In this study, four independent specific natural mutations that cause the loss of both motility and secreted lipase activity were identified in BT2 strains from the United States, United Kingdom, and mainland Europe. Each of these was a unique mutation in either fliR, flhA, or flhB, all of which are genes predicted to encode essential components of the flagellar secretion apparatus. Our results demonstrate the existence of independent mutations leading to the BT2 phenotype; thus, this phenotype has emerged separately at least four times. In addition, BT2 strains from the United Kingdom were shown to have the same mutant allele found in U.S. BT2 strains, suggesting a common origin of this BT2 lineage. This differentiation of distinct BT2 lineages is of critical importance for the development and validation of alternative vaccines or other treatment strategies intended for the control of BT2 strains.

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.

Nucleotides and the adenylate energy charge as indicators of stress in rainbow trout (Oncorhynchus mykiss) subjected to a range of dissolved oxygen concentrations

Abstract Liver nucleotides (ATP, ADP, AMP, IMP), the adenylate energy charge (AEC), total adenylate concentration (TA), and IMP-load were used as measures of stress in rainbow trout (Oncorhynchus mykiss) acclimated to normoxic (10.0 mg/l), hypoxic (6.5 mg/l), and supersaturated (13.0 mg/l) dissolved oxygen concentrations and subjected to a challenge by confinement. Liver ATP (783.0 nmol/g) was significantly different in the normoxic fish compared to either hyperoxic (447.7 nmol/g) or hypoxic (402.0 nmol/g) fish at the end of the confinement. Within 6.0 hr in the confinement, liver AEC in the normoxic fish increased significantly (0.58) compared to hypoxic (0.42) and hyperoxic fish (0.42). Similarly, the IMP-load in normoxic fish (0.16) decreased to near prestress levels by 6.0 hr in confinement compared to either the hypoxic (0.31) or hyperoxic (0.30) fish. Nucleotides in liver were significantly affected by the dissolved oxygen treatments and the confinement stress in contrast to the muscle nucleotides which were not.

Communications: Validation of a Solid-Phase Enzyme Immunoassay Technique for the Measure of Plasma Cortisol in Rainbow Trout

Abstract A kit for a solid-phase enzyme immunoassay (SOPHEIA®) of cortisol in human sera was evaluated and validated for measuring cortisol in plasma of rainbow trout Oncorhynchus mykiss. The accuracy of the SOPHEIA was demonstrated by the recovery of exogenous cortisol concentrations of 25, 50, 100, and 250 ng/mL in charcoal-stripped fish plasma. The amounts (mean ± SE) recovered from triplicate samples were 29.9 ± 2.75, 47.5 ± 3.41, 101.7 ± 12.08, and 232.0 ± 11.06 ng/mL, respectively. The intra- and interassay coefficient of variation (CV = 100 × SD/mean) for cortisol levels in undisturbed fish (26.6 ± 1.18 ng/mL) were 14 and 10%, respectively. The intra- and interassay CV for elevated cortisol levels in stressed fish (330.8 ± 19.90 ng/mL) were 8 and 13%, respectively. Cross-reactivity determined for nine steroids in teleostean fish was negligible. Cortisol concentrations in serial dilutions of pooled fish plasma were parallel to the standard curve. Sensitivity (minimum detection limit) was 3.04 ng/mL....