Antagonistic Interactions and Biofilm Forming Capabilities Among Bacterial Strains Isolated from the Egg Surfaces of Lake Sturgeon (Acipenser fulvescens).

Characterization of interactions within a host-associated microbiome can help elucidate the mechanisms of microbial community formation on hosts and can be used to identify potential probiotics that protect hosts from pathogens. Microbes employ various modes of antagonism when interacting with other members of the community. The formation of biofilm by some strains can be a defense against antimicrobial compounds produced by other taxa. We characterized the magnitude of antagonistic interactions and biofilm formation of 25 phylogenetically diverse taxa that are representative of isolates obtained from egg surfaces of the threatened fish species lake sturgeon (Acipenser fulvescens) at two ecologically relevant temperature regimes. Eight isolates exhibited aggression to at least one other isolate. Pseudomonas sp. C22 was found to be the most aggressive strain, while Flavobacterium spp. were found to be one of the least aggressive and the most susceptible genera. Temperature affected the prevalence and intensity of antagonism. The aggressive strains identified also inhibited growth of known fish pathogens. Biofilm formations were observed for nine isolates and were dependent on temperature and growth medium. The most aggressive of the isolates disrupted biofilm formation of two well-characterized isolates but enhanced biofilm formation of a fish pathogen. Our results revealed the complex nature of interactions among members of an egg associated microbial community yet underscored the potential of specific microbial populations as host probiotics.

Flavobacteria colonizing the early life stages of hatchery-incubated Chinook salmon Oncorhynchus tshawytscha (Walbaum 1792) are markedly diverse.

Flavobacterial diseases are significant impediments to hatchery-based fishery conservation and aquaculture productivity worldwide. Recent studies revealed a multitude of novel flavobacteria within the reproductive fluids and unfertilized eggs of feral Chinook salmon Oncorhynchus tshawytscha broodstock, some of which were associated with systemic disease. Herein, embryonated eggs/fry from these broodstock were assayed for flavobacteria while in incubator stacks in three hatcheries over 2 years, as was the water entering hatchery incubators. Overall, >65% of sampled eggs and 38% of fry were colonized by flavobacteria. One hundred and ninety-one egg and fry-associated flavobacterial isolates were characterized phenotypically and via 16S rRNA gene sequencing and phylogenetic analyses, revealing that the majority fell into 22 clades (i.e., 15 Flavobacterium spp. groups and seven Chryseobacterium spp. groups) that varied in presence by facility. Although some matched previously described fish-pathogenic species, the majority were distinct from all described flavobacteria and likely represent novel species. Of concern, iodophor disinfection at the commonly utilized dose/duration for egg-surface disinfection did not eliminate flavobacteria. Results also implicated maternal routes of infection and source water for some flavobacteria. In total, study findings underscore the complexity of flavobacterial ecology within hatchery environments and highlight the need for improved hatchery biosecurity practices.

Isolation and characterization of Flavobacterium columnare strains infecting fishes inhabiting the Laurentian Great Lakes basin.

Flavobacterium columnare, the aetiological agent of columnaris disease, causes significant losses in fish worldwide. In this study, the prevalence of F. columnare infection was assessed in representative Great Lakes fish species. Over 2000 wild, feral and hatchery-propagated salmonids, percids, centrarchids, esocids and cyprinids were examined for systemic F. columnare infections. Logistic regression analyses showed that the prevalence of F. columnare infection varied temporally and by the sex of the fish, whereby females had significantly higher prevalence of infection. A total of 305 isolates of F. columnare were recovered. Amplification of the near complete 16S rRNA gene from 34 representative isolates and subsequent restriction fragment length polymorphism analyses demonstrated that all belonged to F. columnare genomovar I. Phylogenetic analysis of near complete 16S rRNA gene sequences also placed the isolates in genomovar I, but revealed some intragenomovar heterogeneity. Together, these results suggest that F. columnare genomovar I is widespread in the Great Lakes Basin, where its presence may lead to mortality.

New host record and molecular characterization of Dicauda atherinoidi Hoffman & Walker (Bivalvulida: Myxobolidae): a parasite of the emerald shiner Notropis atherinoides Rafinesque, 1818 and mimic shiner Notropis vollucellus Cope, 1865.

Updated morphological and histopathological descriptions for Dicauda atherinoidi (Bivalvulida:Myxobolidae) and an expanded host range are supplemented with the first molecular data and phylogenetic analyses of the genus. Plasmodia were located on the head, ventrum/body and fins of infected emerald shiner Notropis atherinoides Rafinesque, 1818 and mimic shiner Notropis vollucellus Cope, 1865, a new host species. Myxospores were spherical, ranging 9.3-11.4 µm (10.5 ± 0.4) in length, 9.0-11.0 µm (9.7 ± 0.4) in width and 6.6-7.0 µm (6.8 ± 0.2) thick in sutural view, and possessed 2-3 caudal processes (5.3-68.3 µm, 31.1 ± 13.6) connected to the spore body at the sutural groove, all of which are consistent with the genus Dicauda. In the absence of available Dicauda sequence data, the 18S rDNA sequences from Michigan isolates were most similar to Myxobolus spp. Phylogenetic analyses clustered these isolates with myxobolid species from cyprinid fish, suggesting these parasites may represent an underpopulated group of cyprinid-infecting myxozoans. Histopathology revealed thin-walled plasmodial pseudocysts in the dermis and associated connective tissue, where granulomatous inflammation and focal scale atrophy were also present. Further sampling/sequencing of myxobolids from Notropis spp. should expand these underrepresented myxozoans and offer further insight into Myxobolidae host family tropisms.

Polyphasic characterization of Aeromonas salmonicida isolates recovered from salmonid and non-salmonid fish.

Michigan's fisheries rely primarily upon the hatchery propagation of salmonid fish for release in public waters. One limitation on the success of these efforts is the presence of bacterial pathogens, including Aeromonas salmonicida, the causative agent of furunculosis. This study was undertaken to determine the prevalence of A. salmonicida in Michigan fish, as well as to determine whether biochemical or gene sequence variability exists among Michigan isolates. A total of 2202 wild, feral and hatchery-propagated fish from Michigan were examined for the presence of A. salmonicida. The examined fish included Chinook salmon, Oncorhynchus tshawytscha (Walbaum), coho salmon, O. kisutcha (Walbaum), steelhead trout, O. mykiss (Walbaum), Atlantic salmon, Salmo salar L., brook trout, Salvelinus fontinalis (Mitchill), and yellow perch, Perca flavescens (Mitchill). Among these, 234 fish yielded a brown pigment-producing bacterium that was presumptively identified as A. salmonicida. Further phenotypic and phylogenetic analyses identified representative isolates as Aeromonas salmonicida subsp. salmonicida and revealed some genetic and biochemical variability. Logistic regression analyses showed that infection prevalence varied according to fish species/strain, year and gender, whereby Chinook salmon and females had the highest infection prevalence. Moreover, this pathogen was found in six fish species from eight sites, demonstrating its widespread nature within Michigan.

Bacterial infections of Chinook salmon, Oncorhynchus tshawytscha (Walbaum), returning to gamete collecting weirs in Michigan.

Herein, we describe the prevalence of bacterial infections in Chinook salmon, Oncorhynchus tshawytscha (Walbaum), returning to spawn in two tributaries within the Lake Michigan watershed. Ten bacterial genera, including Renibacterium, Aeromonas, Carnobacterium, Serratia, Proteus, Pseudomonas, Hafnia, Salmonella, Shewanella and Morganella, were detected in the kidneys of Chinook salmon (n = 480) using culture, serological and molecular analyses. Among these, Aeromonas salmonicida was detected at a prevalence of ∼15%. Analyses revealed significant interactions between location/time of collection and gender for these infections, whereby overall infection prevalence increased greatly later in the spawning run and was significantly higher in females. Renibacterium salmoninarum was detected in fish kidneys at an overall prevalence of >25%. Logistic regression analyses revealed that R. salmoninarum prevalence differed significantly by location/time of collection and gender, with a higher likelihood of infection later in the spawning season and in females vs. males. Chi-square analyses quantifying non-independence of infection by multiple pathogens revealed a significant association between R. salmoninarum and motile aeromonad infections. Additionally, greater numbers of fish were found to be co-infected by multiple bacterial species than would be expected by chance alone. The findings of this study suggest a potential synergism between bacteria infecting spawning Chinook salmon.

[Digital transformation in urology-opportunity, risk or necessity?] / Digitale Transformation in der Urologie ­ Chance, Risiko oder Notwendigkeit?

Ultimately, new (digital) techniques and artificial intelligence (AI) applications are changing the working environment in urology. This can be an opportunity for further development, but also a change which is not desired. Adjustments to work processes may be necessary. So-called disruptive processes lead to fundamental changes. In the context of the digital transformation, our way of working is changing. Classic hierarchies, working hours, and working environments are dissolving in favor of creative and flexible working models and corporate structures. Clinics and practices in urology must prepare themselves for changing requirements and be able to provide answers.

[Artificial intelligence : What do urologists need to know?] / Künstliche Intelligenz : Was müssen Urolog*innen wissen?

In the past 10 years, the methods of artificial intelligence (AI) have experienced breakthroughs that have opened up a multitude of new fields of application for information technology. AI is particularly strong in those areas where patterns have to be recognized and conclusions and forecasts based on large, multiparametric data sets have to be drawn. Computers are superior to us in terms of precision and speed in these problems. These advances in information technology reach us at a time when innovations in diagnostics and sensor technology enable more precise patient stratification and confront medical personnel with an increasing quantity and quality of patient data. Urology is symbolic of this new complexity of medicine, in which multi-layered diagnostic cascades require a high degree of interdisciplinarity and, especially in uro-oncology, therapeutic strategies are becoming more differentiated and require the interpretation of multiple clinical and diagnostic data. Here, methods of Artificial Intelligence will in future support medical personnel in diagnostics and therapy decisions and thus come closer to the goal of precision medicine. A prerequisite for the success of AI-based support tools will be the transparent development and validation of the software, as well as the population-based visualization of decision parameters.

[Innovative approaches in prostate cancer ultrasound]. / Innovative Sonographie beim Prostatakarzinom.

Today, systematic random biopsies have virtually replaced ultrasound as an imaging tool in the early diagnosis and staging of prostate cancer. Transrectal ultrasonography (TRUS) is now utilized almost only to guide the biopsy needle into the correct anatomical or topographical region of the prostate. Nevertheless, a large number of clinically significant carcinomas are not discovered despite of multiple systematic biopsies. This has led to a dramatic increase in the number of biopsy samples taken, with 6, 10, 12 to 143 being taken during one session depending on the site. Newer modalities and innovative techniques are being investigated in order to accurately identify patients with prostate cancer at different stages of the disease. Innovative ultrasonography techniques may improve the diagnosis and staging of current imaging techniques.

Direct amifostine effect on renal tubule cells in rats.

Clinical trials indicate that amifostine offers protection against cisplatin-induced nephrotoxicity. It is unclear whether a direct pharmacological t on renal tubular cells is involved. We investigated the effect of amifostine pretreatment on the tubular apparatus and evaluated its nephroprotective potential. A total of 32 rats were treated by i.p. administration of 0.9% saline solution (group 1), 5 mg/kg cisplatin (group 2), 25 mg/kg amifostine (group 3), and 25 mg/kg amifostine followed by 5 mg/kg cisplatin (group 4) after 30 min. We recorded elevation of N-acetyl-beta-D-glucosaminidase (NAG) in 24 h pooled urine as a specific marker for tubular lesions, renal leakage of magnesium as an unspecific nephrotoxicity marker, and survival over a 10-day observation period. A significant (P < 0.002) increase in urinary NAG after treatment was documented only in cisplatin-treated group 2 [day 2 (mean+/-SE), 93+/-2.1 units/gram creatinine; day 4, 70.6+/-16 units/gram creatinine; normalization at day 8]. Treatment with amifostine before cisplatin administration resulted in a slight urinary NAG leakage (day 2, 2.8+/-1.8 units/gram creatinine; day 4, 13.8+/-13 units/gram creatinine; normalization at day 6). No increase in urinary enzyme levels was seen in the other groups, and there were no significant differences in urinary magnesium between all groups. Four of eight rats in the cisplatin-treated group and one of eight rats in the amifostine plus cisplatin-treated group died.

[Prostate biopsy: Procedure in the clinical routine]. / Prostatabiopsie : Durchführung im klinischen Alltag.

Over the last decade there has been a 25% decrease in the mortality rates for prostate cancer. The reasons for this significant decrease are most likely associated with the application of urological screening tests. The main tools for early detection are currently increased public awareness of the disease, prostate-specific antigen (PSA) tests and transrectal ultrasound (TRUS) guided topographically assignable biopsy sampling. Together with the histopathological results these features provide essential information for risk stratification, diagnostics and therapy decisions. The evolution of prostate biopsy techniques as well as the use of PSA testing has led to an increased identification of asymptomatic men, where further clarification is necessary. Significant efforts and increased clinical research focus on determining the appropriate indications for a prostate biopsy and the optimal technique to achieve better detection rates. The most widely used imaging modality for the prostate is TRUS; however, there are no clearly defined standards for the clinical approach for each individual biopsy procedure, dealing with continuous technical optimization and in particular the developments in imaging. In this review the current principles, techniques, new approaches and instrumentation of prostate biopsy imaging control are presented within the framework of the structured educational approach.

Identification of a microsatellite region composed of a long homopurine/homopyrimidine tract surrounded by AT-rich sequences upstream of the rat stress-inducible hsp 70.1 gene.

A DNA region containing several repetitive motifs has been detected about 1.9 kbp upstream of the transcription unit of the rat stress-inducible hsp 70.1 gene. The most interesting element of this area is a microsatellite sequence (GA)6CAG(TC)24 that consists of an inverted repeat partially overlapping with the long homopurine/homopyrimidine tract (Pu/Py). DNA molecule within the described sequence can theoretically adopt alternate, non-B structures (H-DNA or cruciform) containing single-stranded regions. This microsatellite region is flanked by AT-rich sequences containing several poly(A) tracts. The longest of them with a possible potential to destabilized a double-stranded DNA helix is localized around 160 bp downstream the (GA)6CAG(TC)24. The DNA fragment containing sequences described above was subcloned into the pUC19 vector and the resulting plasmid was subjected to the standard S1 susceptibility assay. Preliminary mapping of the S1 cleavage site indicates for the formation of the non-B-DNA structure within the Pu/Py tract. This is to our knowledge a first report on the existence of a complex microsatellite region on upstream the 5'-end of the hsp 70 gene in mammals.

Quantitative RT-PCR assay for mRNA of VEGF and histone H4 in the determination of proliferative and angiogenic activity in vulvar pathology.

Proliferative and angiogenic activity of tissue specimens taken from women with various vulvar pathologies were evaluated by determining the number of mRNA VEGF molecules and H4 histone mRNA molecules, by means of the QRT-PCR (TaqMan) technique. Following a cluster analysis the results, where normalised. Euclidean distances were used, all the cases were classified into three groups of pathologies. Group I included low degree vulvar pathologies, group II included high degree vulvar pathologies and group III included vulvar pathologies with high proliferative and angiogenic activity. Significant differences were found in the proliferative and angiogenic activity between groups I and III, and between groups II and III, while no statistically significant differences were found between groups I and II.

Pneumocystis carinii pneumonia in the Federal Republic of Germany in the era of changing antiretroviral therapy - IDKF 13 -. German AIDS Study Group (GASG/IdKF).

BACKGROUND: Pneumocystis carinii pneumonia (PCP) is one of the most common AIDS defining diagnoses. METHODS: In a prospective observational trial all cases of Pneumocystis carinii pneumonia (PCP) were evaluated. Patients with and without PCP-prophylaxis were compared for symptoms, efficacy, side effects and mortality at week 4 and 26. RESULTS: 293 patients developed a PCP episode. Patients with no prophylaxis had a significant lower CD4 cell count and a more severe clinical status at time of diagnosis. This was pronounced in the group with first positive HIV test at time of diagnosis. There was no difference in the rate of successful treatment between both groups. At week four a tendency to a better survival in the group with prophylaxis was observed, however this changed to a trend to a better survival at week 26 for the group without prophylaxis. CONCLUSION: Even in the era of highly active antiretroviral treatment many patients present with PCP. Nearly 60% of patients presented without antiretroviral treatment or PCP-prophylaxis. Nearly 25% of all patients had their first HIV-test at time of PCP diagnosis.

[Computerized supported transrectal ultrasound (C-TRUS) in the diagnosis of prostate cancer]. / Computergestützter transrektaler Ultraschall (C-TRUS) in der Diagnostik des Prostatakarzinoms.

In the diagnosis of prostate cancer digital rectal examination and transrectal ultrasound (TRUS) are the most utilized methods for clinical evaluation. However, both methods are not able to differentiate between benign and malignant findings with a high amount of certainty. Nevertheless, TRUS is an excellent tool to guide biopsies in practically any region of the prostate. The most significant problem of visual TRUS interpretation is the lack of specificity, especially being an inexperienced user. In order to enhance the diagnostic capabilities of TRUS we developed a computerized analysis of the TRUS signal information (C-TRUS/ANNA), which was validated by the pathohistologic findings of radical prostatectomies. The question was asked: Can C-TRUS detect cancer that has been missed by even multiple systematic biopsies? The entrance criteria was prior negative systematic random biopsies regardless of number of biopsy sessions or number of individual biopsy cores. Five C-TRUS subvisual algorithms were utilized to evaluate the information of the ultrasound signal. The most suspicious regions were marked by C-TRUS and biopsied by guiding a needle into that specific location. In this study 132 with a history of 6-72 negative systematic random biopsies (median: 12 cores) were evaluated by C-TRUS. The PSA ranged from 3.1-36 ng/ml with a median of 9.01 ng/ml. C-TRUS detected in 66 (50%) of these 132 patients cancer by targeted biopsies. In thes 66 men the median number of negative biopsy sessions were two and a median of 12 biopsy cores had been taken. From the literature, we would expect a cancer detection rate in this group with systematic sextant biopsies of about 7%. Only five of the detected carcinomas showed a Gleason Score (GS) of 5, were as 25 had a GS of 6, 22 a GS of 7 and 15 a GS above 7. The results of this prospective clinical trail indicate that C-TRUS is able to identify clinically significant cancers that were missed by even multiple systematic random biopsies. In addition, the concept of searching for strategies that utilize expertise and refinement of imaging modalities is supported rather than just elevating the number of random biopsies (i.e. 141 cores in one session).

[Therapy of prostate abscess by employing pigtail drainage with transrectal ultrasound control]. / Therapie von Prostataabszessen durch Einlage einer Pigtaildrainage unter transrektaler Sonographiekontrolle.

Prostatic abscess is a rare but often severe disease. In most cases diagnosis is difficult. The best diagnostic tool is transrectal ultrasound, which depicts prostatic abscess very clearly. The therapy of choice to date is surgery, usually transurethral resection. At the Department of Urology of the University of Kiel we have employed another form of treatment, namely perineal drainage of the abscess with a pigtail tube under transrectal ultrasound guidance. This method is expected to be particularly advantageous in younger patients, for whom transurethral resection may cause problems. Five patients aged 24-71 years have been treated in this manner so far.

[Improvement of transrectal ultrasound. Artificial neural network analysis (ANNA) in detection and staging of prostatic carcinoma]. / Weiterentwicklung des transrektalen Ultraschalls. Artifizielle neuronale Netzwerkanalyse (ANNA) in der Erkennung und Stadieneinteilung des Prostatakarzinoms.

As a result of the enhanced clinical application of prostate specific antigen (PSA), an increasing number of men are becoming candidates for prostate cancer work-up. A high PSA value over 20 ng/ml is a good indicator of the presence of prostate cancer, but within the range of 4-10 ng/ml, it is rather unreliable. Even more alarming is the fact that prostate cancer has been found in 12-37% of patients with a "normal" PSA value of under 4 ng/ml (Hybritech). While PSA is capable of indicating a statistical risk of prostate cancer in a defined patient population, it is not able to localize cancer within the prostate gland or guide a biopsy needle to a suspicious area. This necessitates an additional effective diagnostic technique that is able to localize or rule out a malignant growth within the prostate. The methods available for the detection of these prostate cancers are digital rectal examination (DRE) and Transrectal ultrasound (TRUS). DRE is not suitable for early detection, as about 70% of the palpable malignancies have already spread beyond the prostate. The classic problem of visual interpretation of TRUS images is that hypoechoic areas suspicious for cancer may be either normal or cancerous histologically. Moreover, about 25% of all cancers have been found to be isoechoic and therefore not distinguishable from normal-appearing areas. None of the current biopsy or imaging techniques are able to cope with this dilemma. Artificial neural networks (ANN) are complex nonlinear computational models, designed much like the neuronal organization of a brain. These networks are able to model complicated biologic relationships without making assumptions based on conventional statistical distributions. Applications in Medicine and Urology have been promising. One example of such an application will be discussed in detail: A new method of Artificial Neural Network Analysis (ANNA) was employed in an attempt to obtain existing subvisual information, other than the gray scale, from conventional TRUS and to improve the accuracy of prostate cancer identification.

[Gemcitabine/cisplatin vs. MVAC. 5 year survival outcome of the phase III study of chemotherapy of advanced urothelial carcinoma in Germany]. / Gemcitabin/Cisplatin vs. MVAC. 5-Jahres-Ergebnisse der Phase-III-Studie zur Chemotherapie des fortgeschrittenen Urothelkarzinoms in Deutschland.

Of 405 patients with stage IV transitional cell carcinoma from an international multicenter phase III trial, 70 were randomized in Germany to receive either gemcitabine/cisplatin or standard MVAC systemic chemotherapy for locally advanced or metastatic urothelial cancer. Overall survival as the primary endpoint of the study was similar in both arms (median survival GC 15.4 months vs MVAC 16.1 months), as were tumor-specific survival and time to progressive disease. In the intent-to-treat analysis, the 5-year overall survival rate was 10% for patients randomized to GC and 18% randomized to MVAC. Tumor overall response rates (GC 54%, MVAC 53%) were similar. The toxic death rate was 0% in the GC arm and 3% (one patient) in the MVAC arm. Significantly more GC than MVAC patients experienced grade 3/4 anemia (GC 52%, MVAC 20%) with significantly more red blood cell transfusions in the GC arm.Significantly more GC than MVAC patients had grade 3/4 thrombocytopenia (GC 54%, MVAC 17%) without grade 3/4 hemorrhage or hematuria in either arm. More MVAC patients experienced grade 3/4 neutropenia (GC 56%, MVAC 61%, p=1.000), neutropenic or leukopenic fever (GC 0%, MVAC 10%, p=0.237), mucositis (GC 0%, MVAC 7%, p=0.495), and alopecia (GC 6%, MVAC 36%, p=0.004). GC represents a reasonable alternative for the palliative treatment of patients with locally advanced and metastatic transitional cell carcinoma. Sustained long-term survival was only found for patients with locally advanced cancer, lymphatic metastases, or solitary distant metastasis but not for visceral metastatic disease.