Application of low-frequency sonophoresis and reduction of antibiotics in the aquatic systems.

A major concern in aquaculture is the use of chemical therapeutics, such as antibiotics, because of their impact on the environment as well as on the fish product. As a potential tool for reducing antibiotic use, we tested the application of low-frequency ultrasound as a method for enhancing antibiotic uptake. Rainbow trout juveniles (Oncorhynchus mykiss) were exposed to two different concentrations of oxytetracycline (OTC), flumequine (FLU) and florfenicol (FLO), administered by bath after the application of ultrasound. After exposure, concentrations of these substances were measured in the liver and blood of treated fish. Results showed that the ultrasound treatment can significantly increase the uptake for all three antibiotics. The uptake of OTC for example, in fish exposed to an OTC concentration of 20 mg L-1 after prior treatment with ultrasound, was similar to the OTC concentrations in their liver and blood to fish exposed to 100 mg L-1 without sonication. For FLU and FLO, the use of ultrasound caused significant differences of uptake in the liver at high antibiotic concentrations. This suggests that the use of ultrasound as a technique to deliver antibiotics to fish can ultimately reduce the amount of antibiotics discharged into the aquatic environment.

Health care utilization and mortality among elderly patients with myelodysplastic syndromes.

BACKGROUND: Mortality in patients with myelodysplastic syndromes (MDS) is high, and patients are likely to require hospitalizations, emergency department (ED) visits, and transfusions. The relationships between these events and the MDS complications of anemia, neutropenia, and thrombocytopenia are not well understood. PATIENTS AND METHODS: A total of 1864 patients registered in the United States' Surveillance Epidemiology and End Results (SEER) program and aged ≥ 66 years old when diagnosed with MDS in 2001 or 2002 were included. Medicare claims were used to identify MDS complications and utilization (hospitalizations, ED visits, and transfusions) until death or the end of 2005. Mortality was based on SEER data. Kaplan-Meier incidence rates were estimated and multivariable Cox models were used to study the association between complications and outcomes. RESULTS: The 3-year incidence of anemia, neutropenia, and thrombocytopenia was 81%, 25%, and 41%, and the incidence of hospitalization, ED visit, and transfusion was 62%, 42%, and 45%, respectively. Median survival time was 22 months. Cytopenia complications were significantly associated with each of these outcomes. CONCLUSIONS: All types of cytopenia are common among patients with MDS and are risk factors for high rates of health care utilization and mortality. Management of the complications of MDS may improve patient outcomes.

Migratory response of European eel (Anguilla anguilla) phagocytes to the eel swimbladder nematode Anguillicola crassus.

The migratory response of peripheral blood granulocytes and monocytes from the European eel Anguilla anguilla (L.) to infective larvae of the swimbladder nematode Anguillicola crassus Kuwahara, Niimi and Hagaki, 1974 was examined by means of light microscopical histology and with an in vitro assay using a modified Boyden chamber. Histological examination of experimentally infected eels revealed that, already 8 days postinfection, an infiltration of inflammatory cells around L3 of A. crassus in the swimbladder tissue can occur. In the Boyden chamber, in presence of infective larvae of A. crassus (L3), neutrophil granulocytes and monocytes showed a higher migration activity than in the absence of L3. In conclusion, infection of European eels with A. crassus leads to an activation of the defence cells resulting in an increased migration activity compared to uninfected eels.

Vaccination of eels (Anguilla japonica and Anguilla anguilla) against Anguillicola crassus with irradiated L3.

The original host of the swimbladder nematode Anguillicola crassus, the Japanese eel (Anguilla japonica) and the recently colonized European eel (Anguilla anguilla) were immunized with 40 irradiated (500 Gy) 3rd-stage larvae (L3) of this parasite and challenged with an infection of 40 normal L3. The immunization induced a significant reduction of the number of adult worms developing from the challenge infection in A. japonica, but not in A. anguilla. The induced resistance (calculated using the relation of the number of adult worms in immunized eels and in non-immunized control eels) in A. japonica was 87.3%+/-30.4%. Following a single infection, the percentage of adult worms found in A. japonica was lower as compared to A. anguilla, and the few adult worms were much smaller, revealing a lower susceptibility of A. japonica to A. crassus in comparison to A. anguilla. Both eel species developed an antibody response against A. crassus, but the level of antibody responses was not positively correlated with the protection against infection, suggesting that the antibody response is not a key element in resistance of eels against A. crassus. This study suggests that the original host of A. crassus is able to mount efficient protective immune responses against its parasite, whereas the newly acquired host seems to lack this ability.

The swimbladder nematode Anguillicola crassus in the European eel Anguilla anguilla and the Japanese eel Anguilla japonica: differences in susceptibility and immunity between a recently colonized host and the original host.

The swimbladder nematode Anguillicola crassus originates from Asia where it is a parasite of the Japanese eel Anguilla japonica. After its introduction to Europe about 25 years ago, the parasite spread rapidly within the indigenous populations of the European eel Anguilla anguilla and subsequently the prevalence and mean intensity appeared to stabilize. Under experimental and aquaculture conditions the naïve new host appears to be more susceptible to A. crassus compared to the original host. Both eel species develop a immune response against A. crassus. The antibody response is well characterized for the European eel, but poorly characterized for the Japanese eel. It remains unclear if antibodies have any protective function against A. crassus. Encapsulation of larvae of A. crassus can be observed in naturally infected European eels. However, encapsulation of larvae following experimental infection has not been detected in European eels, but only in Japanese eels. Reinfection experiments and intraperitoneal injection of A. crassus homogenates failed to demonstrate the development of acquired immunity in European eels. Immunization with irradiated third stage larvae provided preliminary evidence for acquired immunity against A. crassus in the Japanese eel, but not in the European eel.

Anguillicola papernai (Nematoda: Anguillicolidae) and other helminths parasitizing the African longfin eel Anguilla mossambica.

The swim bladder nematode Anguillicola papernai Moravec & Taraschewski, 1988 has been investigated as regards its occurrence in longfin eels Anguilla mossambica (Peters) in rivers in South Africa. A. papernai revealed a prevalence of around 50% and a mean intensity of about 6 adult worms at 1 sampling site but were less abundant in 3 others. Field observations suggest a more narrow habitat preference than that of Anguillicola crassus and a seasonal pattern of abundance. African longfin eels harboured a poor helminth community. In addition to A. papernai, 2 gastro-intestinal nematodes occurred, the stomach worm Heliconema longissimum Ortlepp, 1923 as the dominant species, and the intestinal Paraquimperia africana Moravec, Boomker & Taraschewski, 2000. Experiments were undertaken using European eels Anguilla anguilla (Linnaeus) and copepods as laboratory hosts. The morphology of larvae and adult parasites obtained from these experimental hosts is described. The ultrastructure of adult worms recovered from wild longfin eels was studied. The 'papilla-like excrescences of fibrous structure' on the adult worms' cuticle, as mentioned in the original description, are in fact the attachment points of thick cords of fibers interconnecting the epicuticle with the hypodermis. Such a structure has not yet been described from any other species of Anguillicola Yamaguti, 1935. At present in South Africa, Mozambique and Madagascar attempts are on the way to establish an eel management like in Asia and Europe including eel farming. In this context, care should be taken to prevent the introduction of non-endemic eel parasites into Africa and Madagascar. On the other hand, the future commercial management of African eel species should not lead to the spread of A. papernai or other parasites of African eel species to Europe or elsewhere. In this study A. papernai has been experimentally demonstrated to be capable of reproducing in the European eel and of using European copepods as intermediate hosts.

Differences in susceptibility of the European eel (Anguilla anguilla) and the Japanese eel (Anguilla japonica) to the swim-bladder nematode Anguillicola crassus.

The swim-bladder nematode Anguillicola crassus originates from the Far East where it is a parasite of the Japanese eel (Anguilla japonica). After A. crassus was introduced to Europe, it became a predominant parasite of the European eel (Anguilla anguilla). A study performed with experimentally infected eels (98 days, 23 degrees C) revealed significant differences in the susceptibility of the two eel species to this parasite. The recovery rate of 30 administered infective A. crassus larvae (L3) from A. japonica was less than half of that from A. anguilla (33.2% and 13.8%, respectively). Almost 60% of the worms recovered from A. japonica were found as dead, encapsulated and necrotic larvae in the swimbladder wall. In contrast, no dead larvae were found in A. anguilla. Additionally, the development of the worms was shown to be significantly slower in A. japonica compared with A. anguilla. The lower survival rate of the worms, together with their slower development, resulted in a significantly lower adult worm burden (11 and 428 mg wet weight, respectively) and in a decreased reproductive success in A. japonica compared with A. anguilla. These results demonstrate that the original host, A. japonica, possesses more effective defence mechanisms against A. crassus than does the non-adapted host, A. anguilla.

Individual and combined effects of cadmium and 3,3',4,4',5-pentachlorobiphenyl (PCB 126) on the humoral immune response in European eel (Anguilla anguilla) experimentally infected with larvae of Anguillicola crassus (Nematoda).

The individual and combined effects of cadmium (Cd) and 3,3',4,4',5-pentachlorobiphenyl (PCB 126) on the antibody response of fish against metazoan parasites were tested. Eels experimentally infected with the swim bladder nematode Anguillicola crassus were exposed to sublethal concentrations of Cd and PCB 126. Cd was added to the water resulting in an effective concentration of 21.7 +/- 12.8 microg/l (mean +/- S.D.). PCB 126 was applied orally at a dose of approximately 100 ng PCB 126 per g body weight. At the end of the experiment, 76 days post-infection (p.i.), eels were found to be infected with 2-3 worms. Immunoblot analyses revealed that the body wall of adult worms was the most suitable crude antigen, and was subsequently used for an ELISA to evaluate the immune response of A. anguilla under various conditions. A significant increase of Anguillicola-specific antibodies in the peripheral blood was first detected 61 days p.i., indicating that it was not the invasive larvae but the adult worms which elicit the antibody response. The presence of Cd in the concentrations applied did not appear to modulate the production of antibodies. In contrast, the exposure to PCB 126 resulted in a complete suppression of the antibody response. A similar effect was also found for the combined exposure of the infected eels to Cd and PCB 126. A suppressed immune response, as demonstrated here, may be the reason why hosts exposed to environmental pollution became often much more easily infected than unexposed conspecifics.

Effects of crude extracts of Mucuna pruriens (Fabaceae) and Carica papaya (Caricaceae) against the protozoan fish parasite Ichthyophthirius multifiliis.

The ciliate Ichthyophthirius multifiliis is among the most pathogenic parasites of fish maintained in captivity. In the present study, the effects of the crude methanolic extract of leaves of Mucuna pruriens and the petroleum-ether extract of seeds of Carica papaya against I. multifiliis were investigated under in vivo and in vitro conditions. Goldfish (Carassius auratus auratus) infected with the parasites were immersed for 72 h in baths with M. pruriens extract, and for 96 h in baths with C. papaya extract. There was a 90% reduction in numbers of I. multifiliis on fish after treatment in baths of each plant extract at 200 mg l(-1 )compared to untreated controls. Consequently, parasite-induced fish mortality was reduced significantly. A complete interruption of trophont recruitment was achieved by immersion in the M. pruriens extract. In vitro tests led to a 100% mortality of I. multifiliis in 150 mg/l M. pruriens extract, and in 200 mg/l of C. papaya extract after 6 h. Although the active constituents of the medicinal plant extracts are still unknown, we have demonstrated that they have potential for effective control of I. multifiliis.

Bowel surveillance patterns after a diagnosis of colorectal cancer in Medicare beneficiaries.

BACKGROUND: Postoperative colon surveillance has been recommended for patients with a diagnosis of local/regional colorectal cancer. The extent to which these recommendations are followed in practice is poorly characterized. Patterns of surveillance after surgery for colorectal cancer were determined by using a large population-based database. METHODS: This is a retrospective cohort study with cancer registry data linked to Medicare claims. Identified were 52,283 patients treated for local/regional colorectal cancer between 1986 and 1996, and surveillance patterns through 1998 were determined. Surveillance patterns were analyzed by using survival analysis and by computing the proportion of surviving patients who underwent procedures during 4 time periods after treatment: 2 to 14 months, 15 to 50 months, 51 to 86 months and more than 87 months. RESULTS: Median times to first through fifth surveillance events were 20, 14, 15, 15, and 15 months, respectively. For 17% of the cohort there was no surveillance event. Younger patients were more likely to undergo surveillance. Surveillance patterns were not affected by stage. The proportions of the cohort that underwent no surveillance during the 4 respective time periods were 54%, 52%, 60%, and 69%. The percentages of patients who underwent surveillance annually or more frequently in the latter 3 time periods, respectively, were 19%, 10%, and 5%, or 11% overall, treating the data for the 3 events as a whole. Over the period from 1986 to 1998, the proportion of patients who had no surveillance procedures gradually decreased, whereas the proportion of those who underwent procedures annually or more frequently remained relatively constant. CONCLUSIONS: During the period from 1986 to 1998 there was low utilization of postdiagnosis colon surveillance in a substantial proportion of elderly patients with a diagnosis of local/regional colorectal cancer. Over time there was a trend toward increasing receipt of any surveillance procedures. The percentages of patients undergoing surveillance annually or more frequently did not change between earlier and later periods.

Quality-of-life outcomes after primary androgen deprivation therapy: results from the Prostate Cancer Outcomes Study.

PURPOSE: To compare health-related quality-of-life outcomes after primary androgen deprivation (AD) therapy with orchiectomy versus luteinizing hormone-releasing hormone (LHRH) agonists for patients with prostate cancer. PATIENTS AND METHODS: Men (n = 431) newly diagnosed with all stages of prostate cancer from six geographic regions who participated in the Prostate Cancer Outcomes Study and who received primary AD therapy but no other treatments within 12 months of initial diagnosis were included in a study of health outcomes. Comparisons were statistically adjusted for patient sociodemographic and clinical characteristics, timing of therapy, and use of combined androgen blockade. RESULTS: More than half of the patients receiving primary AD therapy had been initially diagnosed with clinically localized prostate cancer. Among these patients, almost two thirds were at high risk of progression on the basis of prognostic factors. Sexual function outcomes were similar by treatment group both before and after implementation of AD therapy. LHRH patients reported more breast swelling than did orchiectomy patients (24.9% v 9.7%, P <.01). LHRH patients reported more physical discomfort and worry because of cancer or its treatment than did orchiectomy patients. LHRH patients assessed their overall health as fair or poor more frequently than did orchiectomy patients (35.4% v 28.1%, P =.01) and also were less likely to consider themselves free of prostate cancer after treatment. CONCLUSION: Most endocrine-related health outcomes are similar after surgical versus medical primary hormonal therapy. Stage at diagnosis had little effect on outcomes. These results provide representative information comparing surgical and medical AD therapy that may be used by physicians and patients to inform treatment decisions.

Induction of stress by the swimbladder nematode Anguillicola crassus in European eels, Anguilla anguilla, after repeated experimental infection.

The purpose of this study was to determine under laboratory conditions over a period of 311 days if infection with the nematode Anguillicola crassus induces stress in European eels (Anguilla anguilla), and stimulates the endocrine stress axis as measured by serum cortisol levels. Eels were experimentally infected with 3rd-stage larvae (L3) in different doses to simulate natural conditions with varying infection pressures. Blood samples were drawn from the caudal vein every 2 weeks and serum cortisol concentrations were determined by radioimmunoassay (RIA). The results showed that the application of L3 resulted in a significant increase in the cortisol levels. The period of time at which elevated cortisol values were observed was consistent with the time of larval development and the appearance of adult A. crassus. Thus, there is a stress response to the larval and young adult stages, but no chronic response to older adults. Therefore, it is likely that infection of eels under natural conditions especially with a high number of larvae may be a considerable stressor, in combination with different environmental factors like water temperature, pH, oxygen concentration, pollution and interindividual relationships.

The control of prostate-specific antigen expression and gene regulation by pharmacological agents.

Prostate-specific antigen is a serine protease that is a member of the kallikrein family. It is widely used as an indicator of tumor burden and as a surrogate marker for disease progression in men with androgen-independent prostate cancer. It has been shown that the expression and/or secretion of this glycoprotein can be regulated by pharmacological agents. The effects of these agents on PSA may be independent of their effects on cell growth. For example, a pharmacological agent may down-regulate PSA expression/secretion but have no effect on tumor cell growth. In this case, a patient receiving this therapeutic agent might be falsely considered as having a clinical response. Alternatively, an agent might up-regulate PSA expression/secretion and have an inhibitory effect on cell growth. A patient receiving this therapeutic agent might be diagnosed with progressive disease unless an alternative method for assessing tumor burden is used. Thus, when an agent is to be evaluated in a clinical trial utilizing PSA as a marker for disease progression, it is important to prospectively test whether the agent has an effect on PSA expression and/or secretion. In addition, it is equally important to understand how these regulatory effects relate to cell growth. The purpose of this review is to describe several agents that have been tested for their regulatory effects on PSA and to discuss potential mechanisms of by which this regulation may occur. The implications of these findings in the evaluation of new agents in androgen-independent prostate cancer will be considered.

Evaluation of an ELISA and immunoblotting for studying the humoral immune response in Anguillicola crassus infected European eel Anguilla anguilla.

The applicability of an enzyme-linked immunosorbent assay (ELISA) for the detection of anguillicolosis in feral eels was examined using a crude antigen preparation from the body wall of adult Anguillicola crassus. The screening consisted of samples from 100 feral European eels Anguilla anguilla. As a reference the actual status of infection was determined by dissection of the eels' swim-bladders. The ELISA results were compared with a background value calculated from the results obtained from 43 non-infected farm eels. The screened samples had a high prevalence of A. crassus (83 %); however, the specificity and the negative predictive value of the ELISA were low compared to the high positive predictive value. Nonetheless, the reproducibility (precision) of the test was satisfactory, and for the non-infected reference group specificity was 97.7 %. Although the ELISA, as used in the present study, is not applicable for diagnostic purposes, it represents a useful tool for the investigation of the specific humoral immune response of eels against A. crassus under controlled experimental conditions. Immunoblots using crude antigen preparations from different parts of adult A. crassus as well as a crude somatic third-stage (L3) antigen preparation illustrated that only antigens associated with the body wall of adult A. crassus are potentially suitable for diagnostic purposes. Despite the fact that antibodies against Raphidascaris acus cross-reacted with 3 body wall antigens of A. crassus, the most encouraging results were obtained with the antigen preparation from the outer cuticle of adult A. crassus which yielded a conspicuous, broad band at about 100 kDa.

Humoral immune response of European eel Anguilla anguilla experimentally infected with Anguillicola crassus.

A humoral immune response of the European eel Anguilla anguilla elicited by an experimental infection was demonstrated for the first time against the swimbladder nematode Anguillicola crassus. Eels were experimentally infected once or repeatedly and the antibody response was observed over a period of 325 d. Specific antibodies against A. crassus in the peripheral blood of the eels were measured using an ELISA and the immunoblot technique. Anti-A. crassus antibodies were first observed 8 wk post infection, and appeared to be independent of both the number of infective third stage larvae (L3) administered and the frequency of administration. However, individual eels showed great differences in the course of the antibody response. The late appearance of antibodies in the peripheral blood supports the hypothesis that not the invading L3 but rather the adult parasites elicit the production of specific antibodies. A stage-specific antibody response against the L3 was not observed. Main antigens are located in the body wall, especially in the gelatinous outer cuticle, of adult A. crassus.

Richness and diversity of parasite communities in European eels Anguilla anguilla of the River Rhine, Germany, with special reference to helminth parasites.

A total of 121 European eels (Anguilla anguilla) from 2 sampling sites on the River Rhine were investigated in respect of their parasite communities. Special attention was given to the swim bladders, intestines, gills and fins of the fish. Twelve different parasite species were found to live in and on the eels. Data from each sampling site were kept separate. Parasites found in descending order of prevalence were: Anguillicola crassus, Trypanosoma granulosum, Myxobolus sp., Paratenuisentis ambiguus, Pseudodactylogyrus sp., Bothriocephalus claviceps, Myxidium giardi, Pomphorhynchus laevis, Trichodina sp., Raphidascaris acus, Acanthocephalus lucii and Acanthocephalus anguillae. Significantly different prevalences were reported for L3 larvae of A. crassus, adult P. ambiguus, B. claviceps and Myxobolus sp. at the 2 sampling sites. The highest number of parasite species was recorded from the intestine, which contained up to 6 different helminths. The coexistence of the acanthocephalans P. laevis and P. ambiguus, which showed clear patterns of distribution within the intestine of the respective hosts, was reported for the first time. Up to 3 different helminth species were found in the intestine of individual fish. Among those, acanthocephalans were the most prevalent worms with the eel-specific parasite P. ambiguus as the dominant species not only of the intestinal but also of the total component communities. Both infra and component communities exhibited low diversity and were dominated by this single species. The evenness reached only approximately 50% or less and it remained unclear why the helminth communities of the eels from the River Rhine with its huge catchment area exhibit such a low parasite diversity and high dominance.

Development of Anguillicola crassus (Dracunculoidea, Anguillicolidae) in experimentally infected Balearic congers Ariosoma balearicum (Anguilloidea, Congridae).

The development of Anguillicola crassus in experimentally infected Ariosoma balearicum (Anguilloidea, Congridae) kept in seawater was studied in the laboratory. In parallel trials the effect of water salinity on the development of larval A. crassus in European eels Anguilla anguilla was also investigated using eels kept in seawater of a salinity of 34 per thousand. Both eel species were orally inoculated with L3 larvae of A. crassus and then maintained for up to 3 mo at 18 degrees C in seawater. 110 d post infection, no adult but larval (L3 and L4) stages of A. crassus were detected in the swimbladder wall of Balearic congers, although this period of time was sufficient for the parasites to develop to the adult stage in European eel kept in seawater. The results presented suggest that the definitive host specificity of A. crassus comprises species of the family Anguillidae (i.e. the genus Anguilla), but not members of the Congridae. Theoretically however, A. balearicum might serve as a metaparatenic host. Factors determining the definitive host range of A. crassus remain to be elucidated. Water salinity does not seem to act as a factor affecting definitive host specificity once the parasite has become ingested by the eel.

The epidemiology of prostate cancer part I: descriptive epidemiology.

The incidence and mortality of prostate cancer is highly varied among populations and especially among blacks and whites. The incidence rates of all American populations have dramatically changed over the past 25 years. The recent increase in incidence has been attributed to prostate cancer screening. Although the incidence has increased over the past 25 years, the mortality rates although vastly different between populations have remained rather stable within populations. Prostate cancer is still a disease that primarily afflicts older men. The median age at diagnosis is 71 years for whites and 69 years for blacks. More than 80% are over the age of 65 years. Screening for prostate cancer has dramatically increased the number of men with local disease at diagnosis, but it is unclear whether screening and aggressive treatment have caused a decrease in mortality.

The epidemiology of prostate cancer part II: the risk factors.

The epidemiology of prostate cancer gives us some clues that its etiology is likely both environmental and genetic. There is extreme country to country variability in prostate cancer mortality. Countries in which dietary fat intake is greater have been shown to have higher prostate cancer mortality rates leaving some to conclude that dietary fat causes prostate cancer. Migration studies show that men moving from Japan and China adopt increased risks of prostate cancer. Second- and third-generation Japanese Americans and Chinese Americans actually have risks of prostate cancer similar to white American men. This is highly suggestive that prostate cancer has an environmental influence. The differences in black-white mortality and newer data suggesting a higher mortality among Jamaican and Brazilian men of African descent suggest there may be a genetic predisposition to prostate cancer. Some have suggested certain polymorphisms increase prostate cancer risk, whereas others are searching for genetic mutations that may increase prostate cancer risk. Africans may have an increased prevalence of these genetic risk factors. Ultimately, the cause of prostate cancer is likely to be a combination of environmental and genetic factors.

Impact of low water temperature on the development of Anguillicola crassus in the final host Anguilla anguilla.

The effect of low water temperatures on the development and viability of larval and adult Anguillicola crassus (Nematoda) in the final host Anguilla anguilla was studied. European eels were experimentally infected with A. crassus and then maintained for 4 mo at 4, 9, 10, and 19 degrees C. Larval development showed a temperature-dependent pattern and was significantly retarded at low temperatures. Third-stage larvae survived a 4 mo period at 4 degrees C without being affected, although they were not able to invade the swimbladder wall at this temperature. In contrast, adult worms were severely harmed during a 4 mo period at 4 degrees C, as reflected by increased mortality and decreased growth and reproductivity as compared to the worms maintained for the same period at 18 degrees C. Starvation for the eels at 4 mo at 19 degrees C did not affect the development and growth of the nematode. The experimentally obtained results support the hypothesis that the spread of A. crassus in boreal regions, e.g. Northern Europe, is restricted by the natural ambient temperature regimes.