A 2-stage hierarchical interrupted time-series analysis to quantify the long-term effect of subclinical bacterial kidney disease on performance of farmed Atlantic salmon (Salmo salar L.).

Bacterial Kidney Disease (BKD) is an economically significant disease in salmonid aquaculture and commonly requires antibiotic treatments to reduce its impact. Once a pen of fish is diagnosed with BKD, fish are considered chronically infected, potentially until harvest. Although there appears to be little or no evidence to support it, it is often assumed that subclinical infections affect productivity over the long term. We used a 2-stage hierarchical interrupted time series (ITS) analysis in an attempt to quantify the effect of subclinical BKD on mortality, growth, and food conversion ratio (FCR) of Atlantic salmon cultured in marine farms in Atlantic Canada. For all three outcomes, BKD had for some site cycles a positive effect, and for others a negative effect. Overall, the effect of BKD on mortality and growth could not be detected (effect -0.08 ((95% ci: -0.51, 0.35) and 0.00 (-0.02, 0.02)), while a very small effect showing an increase in FCR was detected (0.07 (-0.01, 0.15)). We hypothesized that minimal interference with fish performance may be compatible with the ecology of Renibacterium salmoninarum, the causative agent of BKD. For this organism, vertical transmission is a primary mode of propagation in low-density host populations as found in the wild. Since farms are always adapting and optimizing their farm management of BKD, these constant adjustments may also have negated our ability to detect the effect of many factors contributing to BKD productivity impacts. Hierarchical ITS analysis is considered an appropriate methodology to investigate the complex relationships with productivity measures over time under farming conditions. In the highly innovative salmon aquaculture industry, health records generating data available for time-series analysis is expected to become more accurate and abundant in the future, providing more opportunities for time-series regression studies.

Differences in Rhodococcus equi Infections Based on Immune Status and Antibiotic Susceptibility of Clinical Isolates in a Case Series of 12 Patients and Cases in the Literature.

Rhodococcus equi is an unusual zoonotic pathogen that can cause life-threatening diseases in susceptible hosts. Twelve patients with R. equi infection in Kentucky were compared to 137 cases reported in the literature. Although lungs were the primary sites of infection in immunocompromised patients, extrapulmonary involvement only was more common in immunocompetent patients (P < 0.0001). Mortality in R. equi-infected HIV patients was lower in the HAART era (8%) than in pre-HAART era (56%) (P < 0.0001), suggesting that HAART improves prognosis in these patients. Most (85-100%) of clinical isolates were susceptible to vancomycin, clarithromycin, rifampin, aminoglycosides, ciprofloxacin, and imipenem. Interestingly, there was a marked difference in susceptibility of the isolates to cotrimoxazole between Europe (35/76) and the US (15/15) (P < 0.0001). Empiric treatment of R. equi infection should include a combination of two antibiotics, preferably selected from vancomycin, imipenem, clarithromycin/azithromycin, ciprofloxacin, rifampin, or cotrimoxazole. Local antibiograms should be checked prior to using cotrimoxazole due to developing resistance.

Host resistance to intranasal Acinetobacter baumannii reinfection in mice.

Acinetobacter baumannii is a major causative agent of healthcare-associated infection and develops multidrug resistance rapidly. However, little is known in the host defense mechanisms against this infection. In this study, we examined if mice recovered from a previous intranasal A. baumannii infection (recovered mice) are fully protected against a subsequent reinfection. We found that, despite the presence of specific serum IgG and mucosal IgA responses prior to the reinfection, the recovered mice were only marginally better protected against intranasal challenge with low doses of homologous or heterologous A. baumannii strains than the naïve mice. Post-challenge immune and inflammatory (cells and cytokines) responses were generally comparable between recovered and naïve mice although the recovered mice produced significantly higher amounts of IFN-γ and IL-17 and had higher percentages and numbers of resident lung CD44(hi)CD62L(-)CD4(+) and CD19(+) B lymphocytes. Taken together, our results suggest that mice recovered from a previous A. baumannii infection remain susceptible to reinfection, indicating the complexity of immune protection mechanism for this Gram-negative, multidrug-resistant emerging pathogen.

[Rhodococcus equi infection in AIDS patients: retrospective analysis of 13 patients in Argentina]. / Infección por Rhodococcus equi en pacientes con SIDA: Análisis retrospectivo de 13 pacientes en Argentina.

INTRODUCTION: Rhodococcus equi is a gram positive coccoid rod that causes pulmonary infections in immunosuppressed patients. METHODS: We retrospectively analyzed epidemiological, clinical, microbiological, radiological, and immunological features as well as the outcomes of 13 AIDS patients with R. equi infection. RESULTS: Between January 1994 and December 2012, 13 patients attending the AIDS department of the Infectious Diseases reference hospital in Buenos Aires were diagnosed with R. equi infection. All were men, the median age was 27 years. At the time of diagnosis, the median of CD4+ T cell counts was 11 cells/µl Twelve patients presented pulmonary disease with isolation of the microorganism from sputum or bronchoalveolar lavage; in the other patient the diagnosis was postmortem with positive culture of cerebrospinal fluid. The most frequent clinical manifestations were fever, haemoptysis, and weight loss. The predominant radiological finding was lobe consolidation with cavitation. Nine patients died after a median survival of 5.5 months. In all of them, cultures persisted positive until the last admission. The other 4 patients did continue clinical follow-ups. CONCLUSION: The insidious course of R. equi disease and the difficulties in the isolation of the microorganism contribute to the delay in the diagnosis and to the high mortality rate of this opportunistic infection.

Infección por Rhodococcus equi en pacientes con SIDA: Análisis retrospectivo de 13 pacientes en Argentina / Rhodococcus equi infection in AIDS patients: Retrospective analysis of 13 patients in Argentina

Introduction: Rhodococcus equi is a gram positive coccoid rod that causes pulmonary infections in immunosuppressed patients. Methods: We retrospectively analyzed epidemiological, clinical, microbiological, radiological, and immunological features as well as the outcomes of 13 AIDS patients with R. equi infection. Results: Between January 1994 and December 2012, 13 patients attending the AIDS department of the Infectious Diseases reference hospital in Buenos Aires were diagnosed with R. equi infection. All were men, the median age was 27 years. At the time of diagnosis, the median of CD4+ T cell counts was 11 cells/μl Twelve patients presented pulmonary disease with isolation of the microorganism from sputum or bronchoalveolar lavage; in the other patient the diagnosis was postmortem with positive culture of cerebrospinal fluid. The most frequent clinical manifestations were fever, haemoptysis, and weight loss. The predominant radiological finding was lobe consolidation with cavitation. Nine patients died after a median survival of 5.5 months. In all of them, cultures persisted positive until the last admission. The other 4 patients did continue clinical follow-ups. Conclusion: The insidious course of R. equi disease and the difficulties in the isolation of the microorganism contribute to the delay in the diagnosis and to the high mortality rate of this opportunistic infection.

Introducción: Rhodococcus equi es un cocobacilo grampositivo que provoca compromiso pulmonar en pacientes inmunodeprimidos. Métodos: En el presente trabajo se analizaron de manera retrospectiva los hallazgos epidemiológicos, clínicos, microbiológicos, imagenológicos, inmunológicos y la evolución de 13 pacientes con SIDA y enfermedad por R. equi. Resultados: Entre enero de 1994 y diciembre de 2012, 13 pacientes internados en la División de VIH/SIDA del hospital de referencia para Enfermedades Infecciosas de la ciudad de Buenos Aires egresaron con diagnóstico de enfermedad por R. equi. Todos eran varones y la mediana de edad fue 27 años. La mediana de linfocitos T CD4+ fue de 11 céls/μl Doce pacientes presentaron enfermedad pulmonar con aislamiento del microorganismo del esputo o del lavado bronco-alveolar; en el restante se recibió post mortem el cultivo positivo de líquido cefalorraquídeo. Las manifestaciones clínicas más frecuentes fueron fiebre, hemoptisis y pérdida de peso. La imagen radiológica predominante fue la consolidación con cavitación. Nueve pacientes fallecieron, con una mediana de supervivencia de 5,5 meses. En todos ellos el cultivo persistió positivo hasta la última internación. Los cuatro restantes abandonaron los controles y no pudieron ser evaluados en el tiempo. Conclusión: El curso insidioso de la enfermedad por R. equi y las dificultades en la identificación del microorganismo, contribuyen al retardo en el diagnóstico y a la elevada mortalidad de esta infección oportunista en esta población de pacientes.

The effect of bacterial dose and foal age at challenge on Rhodococcus equi infection.

While Rhodococcus equi remains the most common cause of subacute or chronic granulomatous bronchopneumonia in foals, development of a relevant model to study R. equi infection has proven difficult. The objective of this study was to identify a challenge dose of R. equi that resulted in slow progressive disease, spontaneous regression of lung lesions and age-dependent susceptibility. Foals less than one-week of age were challenged intratracheally using either 10(6), 10(5), 10(4), 10(3) or 10(2) cfu of R. equi. Two doses (10(3) cfu and 10(5) cfu) were used to challenge 2 and 3-week-old, and 3 and 6-week-old foals, respectively. Physical examination, thoracic ultrasound and blood work were performed. Foals were euthanized at the end of the study or when clinical signs of pneumonia developed. All foals were necropsied and their lung lesions scored. Foals challenged with low concentrations of R. equi developed slow progressive pneumonia and approximately 50% of the foals recovered spontaneously. Likewise, macroscopic (>1cm diameter) pyogranulomatous lesions were only observed when low doses of R. equi were used. Clinical pneumonia was not seen after low dose challenge in the 3-week-old foals or in the 6-week-old foals. This study demonstrates that the use of low doses of R. equi to challenge neonatal foals provides an improved model for studying this disease. Furthermore, susceptibility to R. equi infection was shown to diminish early in the foal's life, as has been reported in the field.

Significant morbidity and mortality attributable to rothia mucilaginosa infections in children with hematological malignancies or following hematopoietic stem cell transplantation.

Rothia mucilaginosa is a gram-positive coccus that poses a diagnostic challenge and often requires DNA pyrosequencing for diagnosis as it can be easily mistaken for coagulase-negative staphylococci on initial culture results. While it is often times normal human oral and upper respiratory tract microbiota, it can be a virulent pathogen in immunocompromised patients. Most commonly, it causes bacteremia (catheter and non-catheter related) and meningitis in these patients. Our objective was to report the incidence of R. mucilaginosa infections in neutropenic children with hematological malignancies or following hematopoietic stem cell transplantation at a major children's hospital. We report 11 patients in this cohort who developed clinically significant R. mucilaginosa infections, including three deaths directly attributable to this microorganism. Three patients developed significant neurological involvement, accounting for two of the deaths, and one patient died of disseminated infection. Except for one, all patients had severe neutropenia, central line catheters, and mucosal breakdown at the time of infection. Patients who succumbed never achieved neutrophil recovery. In conclusion, R. mucilaginosa can lead to life-threatening infections in immunocompromised hosts, especially in profoundly neutropenic patients.

Association between radiographic pattern and outcome in foals with pneumonia caused by Rhodococcus equi.

Our objective was to characterize the association between types of radiographic findings and outcome in foals with pneumonia caused by Rhodococcus equi. Admission lateral thoracic radiographs of 62 foals with culture-confirmed R. equi pneumonia were reviewed retrospectively. A scoring system was developed to individually assess the severity of alveolar pattern, interstitial pattern, tracheobronchial lymphadenopathy, pleural effusion, and the number of nodular opacities and cavitary lesions. Individual scores were added to obtain a total radiographic score ranging from 0 (normal) to 22. Forty-three of 62 foals (69%) survived to discharge. The median total radiographic score of nonsurvivors (14; range, 9-16) was significantly (P = 0.007) higher than that of survivors (11; range, 4-15). Foals with a total radiographic score of greater than or equal to 15 were 6.15 times (95% CI: 1.35 to 28.2) less likely to survive than foals with a lower score (P = 0.019). A multivariate logistic regression model was used to identify the potential associations between specific types of radiographic lesions and outcome. The model was statistically significant (P = 0.002) and correctly classified 75.8% of foals. Only severity of alveolar pattern and number of cavitary lesions made statistically significant contributions to the model. There was no significant association between concurrent isolation of other bacteria along with R. equi and the types or severity of radiographic lesions. Based on the results of this study, severity of alveolar pattern and number of cavitary lesions are the radiographic findings significantly associated with a poor outcome in foals with R. equi pneumonia.

The sensor kinase MprB is required for Rhodococcus equi virulence.

Rhodococcus equi is a soil bacterium and, like Mycobacterium tuberculosis, a member of the mycolata. Through possession of a virulence plasmid, it has the ability to infect the alveolar macrophages of foals, resulting in pyogranulomatous bronchopneumonia. The virulence plasmid has an orphan two-component system (TCS) regulatory gene, orf8, mutation of which completely attenuates virulence. This study attempted to find the cognate sensor kinase (SK) of orf8. Annotation of the R. equi strain 103 genome identified 23 TCSs encoded on the chromosome, which were used in a DNA microarray to compare TCS gene transcription in murine macrophage-like cells to growth in vitro. This identified six SKs as significantly up-regulated during growth in macrophages. Mutants of these SKs were constructed and their ability to persist in macrophages was determined with one SK, MprB, found to be required for intracellular survival. The attenuation of the mprB- mutant, and its complementation, was confirmed in a mouse virulence assay. In silico analysis of the R. equi genome sequence identified an MprA binding box motif homologous to that of M. tuberculosis, on mprA, pepD, sigB and sigE. The results of this study also show that R. equi responds to the macrophage environment differently from M. tuberculosis. MprB is the first SK identified as required for R. equi virulence and intracellular survival.

Randomized clinical field trial of a bacterial kidney disease vaccine in Atlantic salmon, Salmo salar L.

A randomized, blinded clinical trial was performed to assess the relative effectiveness of five commercial and one experimental vaccine in a population of farmed fish experiencing a bacterial kidney disease (BKD) outbreak that occurred in one study cage that was part of a larger clinical field trial. A total of 6000 uniquely identified Atlantic salmon S1 presmolts were randomly assigned to vaccine groups in the hatchery and transferred to a commercial marine aquaculture site. Repeated sampling events to evaluate growth, inherent physical conditions and health status were carried out over the entire production cycle. During the second summer at sea, the study cage developed an outbreak of BKD that lasted approximately 240 days. The effectiveness of the selected vaccines was evaluated using survival analysis methods. The sole vaccine group offering protection for BKD was found to significantly decrease the hazard of dying (hazard ratio, HR = 0.68, P = 0.018) during the outbreak, compared to the industry standard, vaccine group. Additionally, during the outbreak, fish with a shortened operculum had a significantly decreased hazard (HR = 0.38, P = 0.033) compared to those fish with a normal operculum, while fish with jaw deformities had a significantly increased hazard (HR = 2.55, P = 0.001) compared to fish with normal jaw status.

Extrapulmonary disorders associated with Rhodococcus equi infection in foals: 150 cases (1987-2007).

OBJECTIVE: To describe frequency, types, and clinical outcomes of extrapulmonary disorders (EPDs) in foals in which Rhodococcus equi infection was diagnosed, and to identify factors determined at the time of admission that differentiated foals that developed EPDs from foals with R equi infection identified only in the lungs. DESIGN: Retrospective case series. ANIMALS: 150 foals aged 3 weeks to 6 months with a diagnosis of R equi infection. PROCEDURES: Medical records were reviewed for information on date of admission, signalment, history, clinical signs, diagnostic testing, treatment, duration of hospitalization, invoice, and outcome. For each EPD identified, further information was collected on the identification, location, treatment, and outcome of the lesion. RESULTS: Of 150 foals with R equi infections, 111 (74%) had at least 1 of 39 EPDs. Survival was significantly higher among foals without EPDs (32/39 [82%]) than among foals with EPDs (48/111 [43%]), but many EPDs were only recognized after death. Risk factors significantly associated with EPDs included referral status, duration of clinical signs prior to admission, leukocytosis, and neutrophilia. Foals with EPDs also had a higher heart rate and BUN concentration than foals without. CONCLUSIONS AND CLINICAL RELEVANCE: Practitioners should recognize that extrapulmonary manifestations of R equi occur with high prevalence affecting diverse organ systems, that multiple systems are generally affected when EPDs occur, and that suspicion of R equi infection should prompt evaluation and monitoring of extrapulmonary sites. Improved recognition of the presence of these disorders will help practitioners to better advise their clients in the treatment and outcome of foals with R equi infections.

Intracranial abscessation as a natural mortality factor for adult male white-tailed deer (Odocoileus virginianus) in Kent County, Maryland, USA.

Intracranial abscessation is a cause of natural mortality among free-ranging white-tailed deer (Odocoileus virginianus) across portions of the United States and Canada. Intracranial abscesses caused by Arcanobacterium pyogenes disproportionately affect adult male white-tailed deer. From 2003-08, we evaluated the occurrence of intracranial abscessation among adult (> or = 2.5 yr) radiocollared male white-tailed deer (n=33) at a large private property in Kent County, Maryland, USA. We documented mortality and necropsied 26 (79%) of the 33 deer. In 2007, we collected swabs from the antler bases and nasopharyngeal membranes of living male white-tailed deer in Maryland, USA (n=9), and Texas, USA (n=10), and from freshly rubbed (n=7) and unrubbed (n=7) trees in Maryland, USA. Swabs were cultured for the presence or absence of A. pyogenes. In Maryland, USA, nine (35%) of the 26 necropsied radiocollared male deer had intracranial abscesses. Five (56%) of nine Maryland, USA, males, and none (0%) of 10 Texas, USA, males cultured positive for A. pyogenes. No rubbed or unrubbed trees at the Maryland site cultured positive for A. pyogenes. The rate of intracranial abscess among adult male white-tailed deer at the Maryland, USA, site (35%) exceeds reported rates for other regions of the United States (9%).

Decreased mortality of Lake Michigan Chinook salmon after bacterial kidney disease challenge: evidence for pathogen-driven selection?

In the late 1960s, Chinook salmon Oncorhynchus tshawytscha from the Green River, Washington, were successfully introduced into Lake Michigan. During spring from 1988 to 1992, large fish die-offs affecting Chinook salmon occurred in the lake. Multiple ecological factors probably contributed to the severity of the fish kills, but the only disease agent found regularly was Renibacterium salmoninarum, the causative agent of bacterial kidney disease. In this study, survival after challenge by R. salmoninarum was compared between two Chinook salmon stocks: a Lake Michigan stock from Wisconsin (WI) and the progenitor stock from the Green River. We found that the WI stock had significantly greater survival than the Green River stock. Next, the WI and Green River stocks were exposed to the marine pathogen Listonella anguillarum (formerly Vibrio anguillarum), one of the causative agents of vibriosis; survival after this challenge was significantly poorer for the WI stock than for the Green River stock. A close genetic relationship between the Green River and WI stocks was confirmed by analyzing 13 microsatellite loci. These results collectively suggest that disease susceptibility of Lake Michigan Chinook salmon has diverged from that of the source population, possibly in response to pathogen-driven selection.

Real-time PCR for quantification of viable Renibacterium salmoninarum in chum salmon Oncorhynchus keta.

Quantification of msa gene mRNA of Renibacterium salmoninarum, the causative agent of bacterial kidney disease (BKD), was investigated using reverse transcription followed by real-time PCR assay on R. salmoninarum in culture, and in experimentally challenged chum salmon Oncorhynchus keta fry kidney tissues (total of 70 samples) after intraperitoneal (i.p.) injection and bath infection. Correlations of msa gene mRNA concentrations with culturable cell concentrations (as colony forming units [CFU]), determined by drop-plate culture method on selective kidney disease medium (SKDM) agar through a 12 wk incubation time, and msa gene DNA concentrations by real-time PCR assay were examined. Furthermore, ovarian fluid samples from wild chum salmon adults with no clinical signs of disease were collected from 8 rivers and from clinically infected kokanee 0. nerka and masu salmon O. masou that were reared in 1 and 2 hatcheries, respectively (total of 414 samples). All samples were examined by nested PCR assay. Then, positive samples were examined by real-time PCR assays for mRNA and DNA; mRNA was detectable at 8 log units (5.0 x 101 to 5.0 x 10(9) copies p11(-1)) with high correlation (R2 = 0.999). The mRNA concentration correlated with CFU in kidney tissue from fish infected by i.p. injection (R2 = 0.924), by bath infection (R2 = 0.502) and in culture (R2 = 0.888). R. salmoninarum was detected and quantified by real-time PCR assay for mRNA in ovarian fluid samples in both subclinically infected chum salmon adults and clinically infected kokanee and masu salmon adults; detection rates ranged from 0 to 44.4% and concentrations ranged from 9.7 x 10(2) to 5.6 x 10(5) copies pl(-1). These results indicate that real-time PCR assay for the mRNA is a rapid, sensitive and reliable method to detect and quantify the viability of R. salmoninarum in kidney and ovarian fluid samples of salmonid fishes with both clinical and subclinical infection of the pathogen.

CpG oligodeoxynucleotides up-regulate antibacterial systems and induce protection against bacterial challenge in rainbow trout (Oncorhynchus mykiss).

The effects of unmethylated CpG oligodeoxynucleotides (ODN) on the mammalian immune system are relatively well studied but much less is known of their effects on the immune systems of different fish species. Here we show that CpG ODNs significantly enhance the survival of rainbow trout (Oncorhynchus mykiss) following bacterial challenge when used both as stand-alone prophylactic agents, or as adjuvants to a commercially available vaccine. They are also capable of increasing serum lysozyme activity in vivo and stimulating the production of chemoattractant factors for rainbow trout head kidney (HK) leucocytes in vitro.

Oral administration of a live attenuated Salmonella vaccine strain expressing the VapA protein induces protection against infection by Rhodococcus equi.

Rhodococcus equi remains one of the most important pathogens of foals and vaccination strategies to prevent rhodococcosis are under increasing investigation. Attenuated Salmonella strains carrying heterologous antigens offer an advantageous alternative to conventional vaccines, especially because they induce mucosal and systemic immunity. In this work, we expressed the VapA antigen from R. equi in a Salmonella enterica Typhimurium strain, which was able to colonize and persist in the lymphoid tissue of BALB/c mice. Two days after being challenged, oral immunized mice presented a 3- to 7-fold increase in R. equi clearance. This was progressively enhanced during infection and, on the 10th day, a CFU value 50-fold lower than that recovered from non-immunized mice was attained. The number of hepatic granulomas was 2 times lower, and leukocyte infiltration was transiently detected in immunized mice, contrasting with the severe inflammation and necrosis presented by non-immunized mice. Infection with 1 x 10(7)R. equi CFU caused 100% mortality in the control groups, while all immunized mice survived. This protection was associated with the detection of high levels of anti-VapA IgG in the serum of the vaccinated mice, predominantly the IgG2a isotype. Our results suggest that attenuated Salmonella encoding VapA may be used in foals to prevent rhodococcosis.

A regulatory effect of the balance between TNF-alpha and IL-6 in the granulomatous and inflammatory response to Rhodococcus aurantiacus infection in mice.

After i.v. inoculation with Rhodococcus aurantiacus, wild-type (WT) mice develop nonnecrotic, epithelioid granulomas. Because a high level of TNF-alpha is observed during the initial phase postinfection, we examined the extent to which TNF-alpha contributes to granulomatous inflammation using TNF-alpha gene-deficient (TNF-alpha(-/-)) mice. Despite a lack of R. aurantiacus proliferation, TNF-alpha(-/-) mice displayed high mortality rates within 5 days postinfection, as well as a high level of IL-6 in their spleens. Histological examination showed an absence of granuloma formation in TNF-alpha(-/-) mice. Pretreatment of TNF-alpha(-/-) mice with rTNF-alpha failed to restore this granuloma formation but accelerated bacterial removal and cellular recruitment. This rTNF-alpha administration also attenuated IL-6 production, resulting in increased survival rates of TNF-alpha(-/-) mice. Heat-killed R. aurantiacus induced in vitro enhanced mRNA expression and production of IL-6 in macrophages and DCs from TNF-alpha(-/-) mice when compared with WT controls, and treatment of TNF-alpha(-/-) mouse cells with rTNF-alpha decreased the IL-6 secretion. Moreover, anti-TNF-alpha or anti-IL-6 treatment increased IL-6 or TNF-alpha production by WT mouse cells, respectively. These data suggest that the production of TNF-alpha and IL-6 can be negatively regulated by each other. Administration of rIFN-gamma to TNF-alpha(-/-) mice caused immature granulomas in livers, and treatment with both rTNF-alpha and rIFN-gamma led to the formation of mature granulomas. Overall, TNF-alpha appears crucial for bacterial clearance, cellular recruitment, and granuloma formation. The balance between TNF-alpha and IL-6 during the early phase of infection controls the development of the inflammatory response to R. aurantiacus infection.

Both msa genes in Renibacterium salmoninarum are needed for full virulence in bacterial kidney disease.

Renibacterium salmoninarum, a gram-positive diplococcobacillus that causes bacterial kidney disease among salmon and trout, has two chromosomal loci encoding the major soluble antigen (msa) gene. Because the MSA protein is widely suspected to be an important virulence factor, we used insertion-duplication mutagenesis to generate disruptions of either the msa1 or msa2 gene. Surprisingly, expression of MSA protein in broth cultures appeared unaffected. However, the virulence of either mutant in juvenile chinook salmon (Oncorhynchus tshawytscha) by intraperitoneal challenge was severely attenuated, suggesting that disruption of the msa1 or msa2 gene affected in vivo expression.

Immunosuppression in progeny of chinook salmon infected with Renibacterium salmoninarum: re-analysis of a brood stock segregation experiment.

Female spawner infection level and temperature variation through rearing are sufficient to explain in-hatchery mortality rates and infection levels and smolt to adult return ratios (SARs) of progeny of Renibacterium salmoninarum infected spring chinook salmon. Data from published reports and manuscripts regarding a 1988 brood stock segregation experiment that held progeny of highly infected female spring chinook salmon spawners separate from progeny of other spawners during 16 mo of hatchery rearing are analyzed to test the hypothesis that immunosuppression could account for differences in survival and infection levels between the 2 segregates. Immunosuppression, caused by the presence of the p57 antigen of R. salmoninarum in sufficient concentration within the salmon egg before spawning, can account for differences in infection levels, mortality rates, and SARs for each hatchery raceway in that study. This immunosuppression may be characterized by immunotolerance, or might only affect cell mediated immunity, which appears the most effective defense mechanism against R. salmoninarum infection, as antibody production can result in tissue damaging antibody-antigen complexes. Low-temperature mediated immunosuppression can account for the nearly identical trajectories of infection and mortality between the 2 segregates during the first 8 mo of hatchery rearing. There is no evidence of widespread vertical infection from spawner to progeny, nor is there evidence that brood stock segregation reduces overall mortality. Rather, the suppression of cell-mediated immune mechanisms may condemn progeny of highly infected female spawners to an almost certain eventual premature demise.

Prognosis and clinical evaluation of infection caused by Rhodococcus equi in HIV-infected patients: a multicenter study of 67 cases.

OBJECTIVE: To assess the clinical characteristics and the factors that influenced the prognosis of patients with HIV and infection caused by Rhodococcus equi. DESIGN: Observational, multicenter study in 29 Spanish general hospitals. SETTING: These hospitals comprised a total of 20,250 beds for acute patients and served a population of 9,716,880 inhabitants. PATIENTS: All patients with HIV and diagnosed R equi infection until September 1998. RESULTS: During the study period, 19,374 cases of AIDS were diagnosed. Sixty-seven patients were included (55 male patients; mean +/- SD age, 31.7 +/- 5.8 years). At the time of diagnosis of R equi infection, the mean CD4+ lymphocyte count was 35/ micro L (range, 1 to 183/ micro L) and the stage of HIV infection was A3 in 10.4% of patients, B3 in 31.3%, C3 in 56.7%, and unknown in 1.5%. R equi was most commonly isolated in sputum (52.2%), blood cultures (50.7%), and samples from bronchoscopy (31.3%). Chest radiographic findings were abnormal in 65 patients (97%). Infiltrates were observed in all of them, with cavitations in 45 patients. The most active antibiotics against the strains isolated were vancomycin, amikacin, rifampicin, imipenem, ciprofloxacin, and erythromycin. After a mean follow-up of 10.7 +/- 12.8 months, 23 patients (34.3%) died due to causes related to R equi infection and 6 other patients showed evidence of progression of the infection. The absence of highly active antiretroviral therapy (HAART) was independently associated with mortality related to R equi infection (relative risk, 53.4; 95% confidence interval, 1.7 to 1,699). Survival of patients treated with HAART was much higher than that of patients who did not receive this therapy. CONCLUSIONS: Infection by R equi is an infrequent, opportunistic complication of HIV infection and occurs during advanced stages of immunodepression. In these patients, it leads to a severe illness that usually causes a bacteremic, cavitary pneumonia, although HAART can improve the prognosis.