A novel sacral neuromodulation protocol is associated with reduction in removal for device infection.

INTRODUCTION AND HYPOTHESIS: Sacral neuromodulation (SNM) has been established as an effective third-line therapy for non-obstructive urinary retention and urinary urgency-frequency syndrome. Device infection, ranging from 2-10%, is a severe complication usually necessitating device explanation. This study sought to demonstrate an infection protocol founded upon established device implantation risk factors and novel approaches to reduce the incidence of device infection, while maintaining good antibiotic stewardship following best practice statements. METHODS: A single-surgeon protocol was enacted from 2013 to 2022. Preoperatively, nasal swabs were cultured from each patient. If positive for methicillin-resistant Staphylococcus aureus or methicillin-sensitive Staphylococcus aureus, preoperative treatment with intranasal mupirocin was employed. Preoperative cefazolin was administered in patients with negative cultures or MSSA-positive. All protocol patients were given chlorhexidine wipes before surgery and prepped with a chlorhexidine scrub followed by alcohol/iodine paint. Post-procedural antibiotics were not given. Pre-protocol patients from 2011 to 2013 served as controls. RESULTS: Pre-protocol (n = 87) patients had a significantly higher rate of device infection compared to protocol patients (n = 444) in both the percentage of patients experiencing device infection (4.6% vs 0.9%, p = 0.01) and percentage of procedures associated with device infection (2.9% vs 0.5%, p < 0.05). A successful culture of the nares was achieved in 91.4% of protocol patients, with 11.6% MRSA-positive. Risk ratio for infection of pre-protocol/protocol patients was 0.19 (0.05-0.77) with odds ratio 5.1 (1.3-20.0). CONCLUSIONS: Utilization of a novel SNM infection protocol tailored to a patient's preoperative MRSA colonization is associated with a reduction in the overall incidence of explant for device infection while avoiding prolonged postoperative antibiotic regimens. CLINICAL TRIAL REGISTRATION: The study was initiated prior to January 18, 2017 and does not meet the definition of an applicable clinical trial (ACT) as defined in section 402 (J) of the US PHS Act.

Increased prevalence and severity of Kudoa thyrsites (Cnidaria: Myxosporea) in Atlantic salmon Salmo salar exposed to deeper seawater.

Kudoa thyrsites is a myxozoan parasite of marine fish with a global distribution. In British Columbia (BC), Canada, severe infections are associated with an economically significant degradation of fillet quality in farmed Atlantic salmon. Exposures to naturally occurring actinospores at a coastal research laboratory were used to test the hypothesis that the prevalence and severity of K. thyrsites infections acquired by exposure of Atlantic salmon to seawater (SW) of various depths are not different. In Expt 1, fish were exposed to SW from 1, 7 or 13 m below the surface. Following exposure to deeper-sourced SW, the prevalence of K. thyrsites, determined from microscopic examination of muscle histology sections, was greater in all 4 trials and the severity of infection was greater in 2 trials. In Expt 2, infections were compared over time among salmon held in tanks supplied with deep-sourced SW (raw or UV-irradiated) or in a surface net-pen. The infection was observed in 35 of 40 fish sampled between 3 and 6 mo after tank exposure to raw SW. Coincidentally, the parasite was observed in 4 of 40 fish maintained in the net-pen. No consistent association of the parasite infection was observed with temperature; however, reduced salinity and solar radiation were not ruled out as factors which may reduce the risk of infection from surface SW. These findings require verification at commercial aquaculture sites in BC, as they will inform considerations related to farm siting and net-pen configuration.

Development of a Piscirickettsia salmonis immersion challenge model to investigate the comparative susceptibility of three salmon species.

Piscirickettsia salmonis, the aetiological agent of salmonid rickettsial septicaemia (SRS), is a global pathogen of wild and cultured marine salmonids. Here, we describe the development and application of a reproducible, standardized immersion challenge model to induce clinical SRS in juvenile pink (Oncorhynchus gorbuscha), Atlantic (Salmo salar) and sockeye salmon (O. nerka). Following a 1-hr immersion in 105 colony-forming units/ml, cumulative mortality in Atlantic salmon was 63.2% while mortality in sockeye salmon was 10%. Prevalence and levels of the bacterium in kidney prior to onset of mortality were lower in sockeye compared with Atlantic or pink salmon. The timing and magnitude of bacterial shedding were estimated from water samples collected during the exposure trials. Shedding was estimated to be 82-fold higher in Atlantic salmon as compared to sockeye salmon and peaked in the Atlantic salmon trial at 36 d post-immersion. These data suggest sockeye salmon are less susceptible to P. salmonis than Atlantic or pink salmon. Finally, skin lesions were observed on infected fish during all trials, often in the absence of detectable infection in kidney. As a result, we hypothesize that skin is the primary point of entry for P. salmonis during the immersion challenge.

Factors associated with severity of naturally occurring piscirickettsiosis in netpen- and tank-reared juvenile Atlantic salmon at a research aquarium in western Canada.

The opportunistic examination of factors associated with an outbreak of piscirickettsiosis (SRS) is described in Atlantic salmon Salmo salar post-smolts held in an open netpen or in tanks supplied with raw sea water at a research aquarium in western Canada. During the outbreak, seawater temperature was significantly higher and salinity significantly lower in the netpen compared with the tanks. Mortality in the netpen began approximately 3 weeks prior to that in the tanks, and cumulative mortality in the netpen (34%) was significantly higher than in the tanks (12%). Piscirickettsia salmonis was confirmed by qPCR in tissues from moribund and dead fish and from colonies grown on enriched blood agar medium. Neither P. salmonis nor SRS were observed in salmon held concurrently in UV-irradiated sea water. The elevated mortality was curtailed by treatment with oxytetracycline. These observations further indicate warmer, less saline and periodically hypoxic seawater are risk factors for SRS. UV irradiation of sea water is shown to be a tool for SRS management in fish-holding facilities.

Host size influences prevalence and severity of Kudoa thyrsites (Cnidaria: Myxosporea) infection in Atlantic salmon Salmo salar.

Kudoa thyrsites is a cosmopolitan myxozoan parasite of marine fish. The infection causes an economically important myoliquefaction in farmed Atlantic salmon in British Columbia, Canada. Laboratory exposure of Atlantic salmon smolts to infectious seawater was used to test the hypothesis that infection with K. thyrsites is more severe in age-matched, smaller salmon. In each of 2 trials approximately 4 mo apart, smolts were graded into small (80 and 68 g), medium (117 and 100 g) and large (142 and 157 g) initial weight groups (IWGs) and concurrently exposed to infectious seawater. The effects of IWG and time on fish size and infection severity were assessed by linear mixed-effects models. The fish were screened for infection by histological examination at intervals following exposure. Increases in mean length and weight were statistically significant in all IWG during both trials. The infection was detected in fish in both trials, and in Trial 2, the prevalence was significantly greater in larger fish 1000 degree-days (DD) after exposure. The severity of infection (plasmodia mm-2 muscle) was significantly higher in larger smolts: between medium and large IWGs at 2500 DD in Trial 1 and between small and medium IWGs at 1500 and 2000 DD in Trial 2. The hypothesis is rejected and possible explanations for the greater occurrence of K. thyrsites in larger smolts are discussed.

Synergistic osmoregulatory dysfunction during salmon lice (Lepeophtheirus salmonis) and infectious hematopoietic necrosis virus co-infection in sockeye salmon (Oncorhynchus nerka) smolts.

While co-infections are common in both wild and cultured fish, knowledge of the interactive effects of multiple pathogens on host physiology, gene expression and immune response is limited. To evaluate the impact of co-infection on host survival, physiology and gene expression, sockeye salmon Oncorhynchus nerka smolts were infected with the salmon louse Lepeophtheirus salmonis (V-/SL+), infectious hematopoietic necrosis virus (IHNV; V+/SL-), both (V+/SL+), or neither (V-/SL-). Survival in the V+/SL+ group was significantly lower than the V-/SL- and V-/SL+ groups (p = 0.024). Co-infected salmon had elevated osmoregulatory indicators and lowered haematocrit values as compared to the uninfected control. Expression of 12 genes associated with the host immune response was analysed in anterior kidney and skin. The only evidence of L. salmonis-induced modulation of the host antiviral response was down-regulation of mhc I although the possibility of modulation cannot be ruled out for mx-1 and rsad2. Co-infection did not influence the expression of genes associated with the host response to L. salmonis. Therefore, we conclude that the reduced survival in co-infected sockeye salmon resulted from the osmoregulatory consequences of the sea lice infections which were amplified due to infection with IHNV.

Differential Effects of Adult Salmon Lice Lepeophtheirus salmonis on Physiological Responses of Sockeye Salmon and Atlantic Salmon.

The salmon louse Lepeophtheirus salmonis, a type of sea lice (family Caligidae), is enzootic in marine waters of British Columbia and poses a health risk to both farmed Atlantic Salmon Salmo salar and wild Pacific salmon Oncorhynchus spp. At the adult stage, sea lice infections can often result in severe cutaneous lesions in their salmonid hosts. To evaluate and compare the physiological consequences of adult L. salmonis infections, smolts of Atlantic Salmon and Sockeye Salmon O. nerka were exposed to 2 (low), 6 (medium), or 10 (high) adult female lice/fish. Mean lice abundance decreased over time in all groups. Skin disruption due to parasite infection was observed in both species. Plasma samples were collected from infected fish and uninfected controls at 1, 3, 5, and 7 d postinfection and measured for indicators of osmoregulatory function and stress. Sockeye Salmon, regardless of L. salmonis exposure level, showed a rapid onset of elevated osmolality and sodium and chloride ion concentrations which were sustained until 7 d postinfection when values returned to levels comparable with the unexposed controls. Conversely, these effects were not measured in Atlantic Salmon. Additionally, differential host effects in blood glucose levels were observed, with Sockeye Salmon displaying immediate elevation in glucose. Relative to Atlantic Salmon, infection with L. salmonis caused a profound physiological impact to Sockeye Salmon characterized by loss of osmoregulatory integrity and a stress response. This work provides the first comprehensive report of the physiological consequences of infections with adult L. salmonis in Sockeye Salmon smolts and helps to further define the mechanisms of susceptibility in this species.

Transmission potential of infectious hematopoietic necrosis virus in APEX-IHN®-vaccinated Atlantic salmon.

Infectious hematopoietic necrosis virus (IHNV) outbreaks have had a significant negative impact on Atlantic salmon Salmo salar production in British Columbia, Canada, since the first outbreak was reported in 1992. In 2005, the APEX-IHN® vaccine was approved for use in Canada for prevention of IHN. The vaccine was proven to be safe and efficacious prior to approval; however, it is unknown as to whether APEX-IHN®-vaccinated Atlantic salmon infected with IHNV can support replication and virus shedding in sufficient quantities to provide an infectious dose to a nearby susceptible host. To determine whether vaccinated, infected fish are able to transmit an infectious dose of IHNV, vaccinated Atlantic salmon were injected with IHNV (104 plaque-forming units per fish) and cohabitated with either naïve Atlantic salmon or naïve sockeye salmon Oncorhynchus nerka. APEX-IHN®-vaccinated fish were significantly protected against IHNV with mortality occurring in only 2.6% of the population as opposed to 97% in unvaccinated controls. Vaccination in IHNV-infected Atlantic salmon completely abolished disease transmission to cohabitating naïve sockeye salmon and reduced virus spread among cohabitating naïve Atlantic salmon. At 7 mo post-vaccination, IHNV-neutralizing antibodies were detected in nearly all vaccinated fish (94%) with similar titer occurring between vaccinated, infected fish and vaccinated, uninfected fish, indicating APEX-IHN® vaccination induces a robust seroconversion response. Taken together, these results demonstrate that vaccination greatly reduces the infectious load and potential for IHNV transmission. As such, APEX-IHN® should be included in fish health management strategies when culturing Atlantic salmon in IHNV endemic areas.

Comparison of quantitative PCR and ELISA for detection and quantification of Flavobacterium psychrophilum in salmonid broodstock.

A quantitative PCR (qPCR) assay was developed for Flavobacterium psychrophilum, the causative agent of bacterial coldwater disease. The assay was targeted to fp1493 as it encodes a putative outer membrane protein (FP1493) that is reactive to the monoclonal antibody (MAb FL43) used in a standardized F. psychrophilum capture enzyme-linked immunosorbent assay (ELISA). The qPCR was specific to F. psychrophilum and was able to detect between 8 and 809000 copies of fp1493. To determine if antigen level in the tissue was indicative of bacterial concentration, kidney samples from 108 steelhead Oncorhynchus mykiss and coho salmon O. kisutch female broodstock were screened by ELISA and qPCR. There was no correlation between ELISA optical density (OD) values and the number of F. psychrophilum cells g⁻¹ of kidney tissue as estimated by qPCR (rS = 0.42; p > 0.05). The median number of F. psychrophilum cells in steelhead samples was 6.11 × 10³ cells g⁻¹ of tissue. For coho salmon samples, the median number of cells was 3.95 × 10³ cells g⁻¹ of tissue. Agreement between the 2 assays was less than 50%. As fp1493 is a single-copy gene and differential expression of FP1493 has been reported, we hypothesize that the discrepancy between the 2 assays is due to increased expression of FP1493 in the in vivo environment. Therefore, ELISA OD values most likely provide an indication of differential protein expression, while the qPCR assay estimates bacterial load in tissue.

Enhanced efficacy of an attenuated Flavobacterium psychrophilum strain cultured under iron-limited conditions.

An attenuated strain of Flavobacterium psychrophilum (CSF259-93B.17) has shown potential as a vaccine for prevention of bacterial coldwater disease (BCWD) in rainbow trout, Oncorhynchus mykiss (Walbaum). Because BCWD outbreaks can result in high mortality in other salmonid species, specifically coho salmon, Oncorhynchus kisutch (Walbaum), the live-attenuated strain was tested as a vaccine in this species. Additionally, we hypothesized that culture of the vaccine strain under iron-limited conditions would lead to improved protection against BCWD. To test this hypothesis, coho salmon were either injection or immersion immunized with CSF259-93B.17 cultured in iron-replete or iron-limited medium. Resultant antibody titers were low and not significantly different between the two treatments regardless of vaccine delivery method (P > 0.05). Following injection challenge with a virulent F. psychrophilum strain, mortality for injection vaccinated fish was significantly reduced compared to the control but did not differ by treatment (P > 0.05). Relative percent survival (RPS) was high in both treatments (90% in iron-replete, 98% in iron-limited medium). Fish immunized by immersion with CSF259-93B.17 grown in iron-replete medium exhibited lower mortality (29.3%; RPS 46%) when compared to mock immunized fish, but this was not significant. However, mortality was significantly lower in fish immunized with CSF259-93B.17 grown in iron-limited medium (14.7%; RPS 73%) when compared to mock immunized fish. The results demonstrate that the live-attenuated F. psychrophilum strain can confer protection to coho salmon and vaccine efficacy is enhanced by culturing the strain under iron-limited conditions.

Plasma carotenoid concentrations of infants are increased by feeding a milk-based infant formula supplemented with carotenoids.

BACKGROUND: Human milk is the gold standard of infant nutrition and is a source of important substances, including carotenoids. Infant formulas are designed to mimic the composition and/or performance of human milk, although currently carotenoids are not routinely added to US infant formulas. The aim of this study was to assess plasma concentrations of ß-carotene, lutein and lycopene 56 days after feeding infants milk-based infant formula without (CTRL) or with different concentrations of added carotenoids (L1 and L2). RESULTS: Plasma carotenoid concentrations increased in infants fed carotenoid-supplemented formulas as compared with the control formula with no added carotenoids. At study day 56, infants fed the supplemented formulas (L1 and L2) had mean plasma lutein, ß-carotene and lycopene concentrations that were within the range of a concurrent group of human milk-fed infants (HM). Anthropometric measurements were comparable among all study groups. CONCLUSION: Plasma carotenoid concentrations of infants fed the supplemented formulas were within the range of the HM group and are consistent with reported plasma carotenoid ranges in human milk-fed infants. The experimental formulas were well tolerated and anthropometric measurements were comparable among all study groups.

Pneumonia caused by toxic epidermal necrolysis.

A well-functioning 68 year old gentleman presented to our hospital with a macular rash 2 weeks after starting a course of Ciprofloxacin. There was rapid progression of skin involvement including the mucosa, complicated by pancytopaenia. Toxic Epidermal Necrolysis (TEN) was suspected and the patient was administered intravenous immunoglobulins and granulocyte colony stimulating factor. TEN was confirmed on skin biopsy and a lymphocyte transformation test demonstrated sensitisation to Ciprofloxacin. The patient developed multifocal pulmonary infiltrates with evidence of pulmonary involvement and probable pneumonia after 1 week and was treated with broad spectrum antibiotics. He also became dysphagic and suffered recurrent aspiration pneumonias. Follow up studies revealed fixed airways obstruction and features of bronchiolitis on computed tomography. This case highlights pulmonary involvement which can become a chronic complication of TEN, itself precipitated by the rare drug cause of Ciprofloxacin.

Piscirickettsia salmonis shedding and tissue burden, and hematological responses during cohabitation infections in chum Oncorhynchus keta, pink O. gorbuscha and Atlantic salmon Salmo salar.

BACKGROUND: Salmonid rickettsial septicemia is an emergent and geographically widespread disease of marine-farmed salmonids caused by infection with the water-borne bacterium Piscirickettsia salmonis. Very little is known about the route, timing, or magnitude of bacterial shedding from infected fish. METHODOLOGY/PRINCIPAL FINDINGS: A cohabitation challenge model was used to assess shedding from chum Oncorhynchus keta, pink O. gorbuscha and Atlantic salmon Salmo salar. Infections in donor fish were established by intraperitoneal injection of P. salmonis. Naïve recipients were cohabitated with donor fish after which cumulative percent morbidity and mortality (CMM) was monitored, and bacterial burdens in kidney and in tank water were measured by qPCR. All donor fish died with mean days-to-death (MDD) among species ranging from 17.5 to 23.9. Among recipients, CMM ranged from 42.7% to 77.8% and MDD ranged from 49.7 to 56.4. In each trial, two peaks of bacterial DNA concentrations in tank water closely aligned with the MDD values of donor and recipient fish. Bacterial tissue burden and shedding rate, and plasma physiological parameters were obtained from individual donors and recipients. Statistically significant positive correlations between the shedding rate and P. salmonis kidney burden were measured in donor pink and in donor and recipient chum salmon, but not in donor or recipient Atlantic salmon. In Atlantic salmon, there was a negative correlation between kidney bacterial burden and hematocrit, plasma Ca++ and Mg++ values, whereas in infected chum salmon the correlation was positive for Na+ and Cl- and negative for glucose. CONCLUSIONS: A dependency of bacterial shedding on species-specific patterns of pathogenesis was suggested. The coincidence of bacterial shedding with mortality will inform pathogen transmission models.