The occurrence of Glugea stephani (Hagenmuller, 1899) in American winter flounder, Pseudopleuronects americanus (Walbaum) from the New York-New Jersey lower bay complex

American winter flounder, Pseudopleuronectes americanus , Collected in the New York-New Jersey bay complex, contained cysts in the intestinal tract which were subsequently identified as the microsporidan Glugea stephani . These fish represent 13 collections taken at approximately monthly intervals over a one and a half year period. The winter flounder varied in size from 8 to 33 cm and consequently represented a fish population of diverse age. During this study, a total of 1840 apparently healthy fish were dissected and examined for the presence of Glugea cysts , with an overall parasite infection rate of 6.63%. There was no correlation between size of the fish and percentage of infection. This percentage represents the mean carrying rate of this parasite in the fish population, since these are the fish that have survived the initial infection. This report extends the reported host range to the New York-New Jersey area. As a further aid in the characterization of G. stephani , we have included ultrastructural details of the spore morphology.     

Effect, distribution, and prevalence of Glugea stephani (Microspora) in winter flounder (Pleuronectes americanus) living near two pulp and paper mills in Newfoundland

A study was conducted to determine the effects, geographical distribution, and prevalence of a microsporan parasite, Glugea stephani, in winter flounder (Pleuronectes americanus) in Newfoundland. Fish were captured by SCUBA divers in several coastal areas, including 2 embayments where pulp and paper mill effluent was discharged, as well as a number of pristine sites. Fish health was assessed by comparing histological profiles, condition factors (K), organosomatic indices and blood values between infected and uninfected samples. Multifocal xenomas of G. stephani were observed in several organs of fish taken near contaminated sites, whereas infected samples captured at a pristine site harbored the cysts only in the wall of the digestive tract. Proliferative inflammation, granuloma formation, and focal necrosis were associated with the infection primarily in the liver and kidney. Condition factors and blood values were lower and ovarian development inhibited or delayed in infected flounder. The multifocal infection occurred only in flounder in 2 embayments in western Newfoundland where pulp and paper mill effluent was discharged. Prevalence varied seasonally, with a peak in autumn and a low in spring. It is likely that the multifocal infection was associated with immunodepression after exposure to the contaminant.     

Experimental infection of American winter flounder (Pseudopleuronectes americanus) with Glugea stephani (Microsporida)

Young-of-the-year Pseudopleuronectes americanus were captured in Sandy Hook Bay, New Jersey and maintained in aquaria at 16–19° C for 115 days in 1982 and 127 days in 1983. Half of them were experimentally exposed to Glugea stephani and the others were sham exposed. During the course of these experiments fish from both groups died as a result of G. stephani infection, demonstrating Glwgea -induced mortality in this species. Infections were found in both control and experimentally exposed fish. Mortality of Glwgea -infected fish in the exposed group was 63% while that in the controls was 28.5%.     

The immunomodulating effect of the microsporidan Glugea stephani on the humoral response and immunoglobulin levels in winter flounder, Pseudopleuronectes americanus

A quantitative study of serum immunoglobulin levels and haemagglutination titres has shown that an injection of the microsporidan Glugea stephani caused a decrease in the humoral response of the winter flounder. These examples demonstrate the effect of temperature on the decrease in total IgM levels and haemagglutination titres in winter flounder simultaneously injected with either G. stephani or horse red blood cells.
Transfer of serum from fish injected with spores 3 weeks earlier may also implicate soluble serum factors, such as prostaglandins or leukotrienes, causing the observed decrease in serum IgM levels.     

Laboratory and field studies on Glugea stephani (Hagenmuller), a microsporidan parasite of pleuronectid flatfishes.

The microsporidan Glugea stephani is a common parasite of juvenile English sole (Parophrys vetulus) in Yaquina Bay, Oregon. Field observations indicated that fish became infected only in the upper estuary where summer temperatures were above 15C and the incidence of infection reached 79.8% in the late fall. Laboratory infections developed and parasite growth occurred only at or above 15C. The parasite was successfully transmitted to juvenile English sole by brine shrimp (Artemia salina) and amphipod (Corophium spinocorne) vectors as well as by direct ingestion of spores by the host. Infections that resulted from ingestion of spore-carrying vectors were much heavier than those resulting from the direct ingestion of spores. The speckled sanddab (Citharichthys stigmaeus), a nonpleuronectid flatfish, and chum salmon (Oncorhynchus keta) were refractory to G. stephani infection in the laboratory.     

Infection of plaice Pleuronectes platessa L. with Glugea (Nosema) stephani (Hagenmüller 1899) (Protozoa: Microsporidia) in a fish farm and under experimental conditions

One tank of plaice in the W.F.A. Fish Cultivation Unit at Hunterston, known to habour an infection with the microsporidian Glugea stephani , was sampled over a 2 year period and showed a mean level of infection of 49.4 % with an apparent trend towards a decrease. Experimental oral transmission was achieved, without the use of intermediate or carrier hosts, and the dependence of G. stephani on high temperatures was demonstrated. Most fish were able to tolerate the infection, but some mortalities in young fish were attributed to the parasite, particularly in the 2 months following the experimental infection. The parasite in the fish farm is believed to have been introduced with plaice stocks imported from more southern waters.     

Altered behaviour and reduced survival of juvenile olive flounder, Paralichthys olivaceus, infected by an invasive monogenean, Neoheterobothrium hirame

Neoheterobothriumhirame is a blood feeding monogenean of olive flounder Paralichthys olivaceus. The parasite was first reported in the mid-1990s from the Sea of Japan and became epidemic within cultured and wild flounder populations after several years. Infected fish often suffer from severe anaemia and thus the parasite is thought to have played an important role in the recent depletion of flounder populations in some areas of Japan. However, the causal mechanism underlying the parasite epidemic and decreases in host populations is unclear because apparently N. hirame infection is not fatal to the host. Here, we tested the hypothesis that N. hirame indirectly reduces the survival of wild juvenile flounder by altering their behaviour and making them more susceptible to predation. We conducted a series of experiments to compare behaviours and predation susceptibility between experimentally infected juvenile P. olivaceus and uninfected fish. Results showed that N. hirame infection increases the activity level, alters diel activity and has negative effects on burrowing performance and swimming endurance. When juvenile flounder cohabitated with predators, the survival rate of infected juveniles was approximately 25% less than that of uninfected fish. We believe this is the first empirical evidence linking N. hirame infection to death of the host through predation. Consequences of N. hirame-induced behavioural change for the survival of juvenile flounder in the wild are discussed. We conclude that recent outbreaks of N. hirame are likely to have been a key factor in the decline of flounder populations in Japan.     

Prevalence and intensity ofCapillaria catostomi (Nematoda: Trichuroidea) in white sucker (Catostomus commersoni) in southern Lake Huron,Canada

Synopsis A total of 314 white suckers (Catostomus commersoni) taken monthly during May to December 1975 and in April and May 1976 from southern Lake Huron, Ontario, Canada, were examined forCapillaria catostomi.Prevalence determined from fresh white suckers (66%) was significantly lower than from frozen white suckers (81%). However, there was no significant difference in prevalence ofC. catostomi in samples taken at monthly intervals; in male and female white suckers; or in the age categories.Mean intensity of infection was not significantly different in fresh (9.6 [± 13.41]) or frozen (10.4 [± 13.11]) white suckers. There was no significant difference in intensities between monthly samples in male and female white suckers or in the age categories. Suckers were infected with 1–172 worms. The percentage of male, female and immature worms did not change with sex or age of host or by month. The frequency distribution showed the parasites were overdispersed.A possible life cycle forC. catostomi involving an oligochaete intermediate host is discussed and an attempt made to explain the almost constant levels of prevalence and intensity.     

Growth, reproduction and recruitment patterns of the wide-eyed flounder, Bothus podas Delaroche (Pisces: Bothidae), from the Azores

The wide-eyed flounder, Bothus podas, occurs in many East Atlantic sub-tropical and temperate inshore areas and is the dominant flatfish in shallow-water areas around the Azores. The aim of this study was to examine the growth, reproductive biology and recruitment patterns of the wide-eyed flounder in the Azores. Adult fish samples were obtained by spear fishing and SCUBA hand netting at Faial Island. Juveniles were obtained from a complementary monthly beach-seining programme undertaken at Porto Pim Bay, Faial. Sex and development stage were determined by macroscopic examination of gonads, whereas age estimates were obtained by counting the number of opaque bands in the otoliths. In the Azores, B. podas reaches 6 years old, which is more than reported for the central Mediterranean, the only other population of this species that has been studied. The studied population has a sex ratio biased to males, which contrasts to a previous observational study, which noted a female-biased sex ratio. Spawning time, as determined from the gonadosomatic ratio and the macroscopic examination of gonads, was between July and September. This time period agrees with behavioural observations in the same area. Sexual maturity was estimated at a total length (TL₅₀) of 14.1 cm and at an age (A₅₀) of 1.7 years. Small juveniles occur on sandy inshore nursery areas for most of the year, but mainly between July and August each year. The presence of small fish throughout most of the year could suggest a protracted settlement period or slow growth on occasions on the shallow-water nursery grounds.     

Patterns in abundance and distribution of juvenile flounder, Platichthys flesus, in Minho estuary (NW Iberian Peninsula)

For 2 years, the flatfish community of Minho estuary (Northwest of Iberian Peninsula) was surveyed monthly with a beam trawl. Flounder, Platichthys flesus, was the most representative species contributing the most (74%) to the total flatfish abundance and was the only species occurring throughout the sampling period and along the entire salinity gradient. The majority of flounder sampled consisted of 0- and 1-year group individuals. In both years, a peak in abundance was recorded in late spring–summer months (from May to August) which coincided with seasonal settlement events. Higher densities of newly-settled flounder were obtained in the most upstream site, located in the tidal freshwater wetland, indicating that this area is an important nursery area. Of the environmental variables tested, temperature and salinity were the best predictors for 0-year group abundance explaining around 30% of the variability, whereas sediment was also important when considering all age groups together. This study supports the generally prevalent idea that estuaries act as nurseries for the early life stages of flounder, with particular relevance to the tidal freshwater estuarine wetlands.     

Plagiorchis muris infection in Apodemus agrarius from northern Gyeonggi-do (Province) near the demilitarized zone

The small intestines of 6 species of rodents and 1 species of insectivore were examined seasonally for Plagiorchis muris infection in 3 different localities in northern Gyeonggi-do (Province), near the demilitarized zone (DMZ). A total of 1,496 animals, including 1,366 Apodemus agrarius, 54 Crocidura lasiura (insectivore), 32 Mus musculus, 28 Micronytus fortis, 9 Eothenomys regulus, 6 Micronys minutus, and 3 Cricetulus triton, were live-trapped at Yeoncheon-gun (n = 351), Paju-shi (804) and Pocheon-gun (343) at 3-mo intervals from December 2004 to September 2005. A total of 1,647 P. muris were collected from 72 (5.3%) A. agrarius. The infection rate was the highest in Pocheon-gun (8.2%), followed by Yeoncheon-gun (5.0%) and Paju-shi (4.2%). A higher infection rate was observed in A. agrarius captured during September (19.4%) than those captured during December (3.0%), June (2.6%), or April (0%). However, the worm burden was the highest in June (av. 32.1/animal), followed by September (24.7), December (4.0), and April (0). None of the other animal species were found infected with P. muris. The results reveal that A. agrarius is a natural definitive host for P. muris, and infection rates and worm burdens vary seasonally and geographically.     

Ground-foraging ants (Hymenoptera: Formicidae) and rainfall effect on pitfall trapping in a deciduous thorn woodland (caatinga), Northeastern Brazil

The semi-arid Caatinga is the fourth largest biome of Brazil, which biota still remains one of the most poorly known, especially with regard to invertebrate groups. In this study, a ground-foraging ant assemblage was surveyed during one year and the effect of rainfall on pitfall trapping was assessed. The study was performed in an area located in the municipality of Pentecoste (3 degrees 48' S - 39 degrees 20' W), in the State of Ceará. A 200m transect with 20 equidistant sampling points was established. Transect sampling was performed once a month during 12 months, over the period August 2008-August 2009. At each sampling point, a pitfall trap partially filled with a mixture of ethanol and monoethylene glycol was placed at the beginning of each month and remained in the field for seven days. 39 species belonging to six subfamilies and 19 genera, plus two unidentified species, were collected, with Pheidole (10 spp.) and Camponotus (8 spp.) being the taxa with the most species. 23 species were frequent, being found in more than 50% of the 12 transect samplings. Five species had an intermediate frequency (25 to 50%), while 13 were relatively infrequent (less than 25%). Most of the species (22) showed low occurrence, being found in less than 10% of the 240 samples (20 samples each month, during 12 months). Only five species were collected in more than 50% of the samples, those species being also responsible for most of the total abundance (number of captured individuals of all species) observed each month. The species-accumulation curves (observed and estimated) indicated that sampling sufficiency was attained, and that about 92% of the estimated ground-foraging ant fauna had been collected. 40 and 29 species were collected in the dry and rainy season, respectively, with monthly species richness ranging from 13 to 28. The total ant abundance showed a drastic decrease during the rainy season, and a negative linear correlation was found between rainfall and total ant abundance (R2 = 0.68). A similar negative linear correlation was found for species occurrences against rainfall (R2 = 0.71), and for mean number of species per pitfall trap against rainfall (R2 = 0.71). However, some species showed equal abundance, occurrence and mean number of individuals per pitfall trap in both seasons, while others showed a much higher abundance and occurrence during the rainy season. Pitfall trapping as a method to sample ground-foraging ant assemblage of the Caatinga biome and potential factors responsible for lower pitfall trap performance during rainy season are discussed.     

Eelgrass,Zostera marina,as essential fish habitat for young‐of‐the‐year winter flounder,Pseudopleuronectes americanus (Walbaum, 1792) in Maine estuaries

Distribution and density by habitat for age‐0, young‐of‐the‐year (YOY) winter flounder, Pseudopleuronectes americanus (Walbaum, 1792), were compared for two Maine estuaries to help define essential fish habitat for this life history stage. Two estuaries (Weskeag River and Penobscot Bay) along Mid‐coast Maine were sampled monthly with daytime 1.0 m² fixed‐frame throw traps around neap low tide, May–December over two consecutive years (2003–2004). Both eelgrass and adjacent sand/mud (20–60 cm deep) were randomly sampled with equal effort (4–12 samples per month) at two sites in both the Weskeag River and Penobscot Bay. Significantly higher densities of YOY winter flounder (2–9 cm TL) occurred in eelgrass relative to sand/mud. Density increased significantly in both habitats in 2004, and was higher in Penobscot Bay relative to the Weskeag River. YOY densities compared by eelgrass coverage within throw traps were found to be significantly higher in eelgrass that exceeded 30% coverage when compared with adjacent sand/mud areas and eelgrass coverage of 10–20%. YOY occurred in all months sampled (May–December); no density differences existed by month. These results indicate that very shallow (<0.6 m) eelgrass habitat is of key importance to YOY winter flounder in Maine estuaries and should be viewed as essential fish habitat (EFH) for this species and life stage.     

Parasitism of Cichlasoma paranaense Kullander, 1983 and Gymnotus carapo Linnaeus, 1814 by Clinostomum complanatum (Rudolphi, 1814) metacercariae in the Taquari river.

The infection, known as yellow spot disease, produced by metacercariae of Clinostomum spp. was studied in fishes of the Taquari river, located in Jataizinho, Paraná State, Brazil. A total of 1,582 specimens, belonging to 36 species, were collected between March 1999 and April 2001. Yellow spot disease was observed only in Gymnotus carapo Linnaeus, 1814 (Gymnotiformes, Gymnotidae) and Cichlasoma paranaense Kullander, 1983 (Perciformes, Cichlidae). This parasitism was generated by metacercariae of Clinostomum complanatum (Rudolphi, 1814) (Digenea, Clinostomidae). Among the 88 specimens of G. carapo examined, 7 (prevalence = 8%) had cysts of the parasite. Four of them were captured in July 1999 and three in October 1999. In the other months, no specimen found was infected. Using relative condition factor (Kn) analysis, it was determined that both infected and non-infected specimens had a total weight equal to the theoretically expected value for each total length (Kn = 1.0). Among 56 individuals of Cichlasoma paranaense, 6 (prevalence = 10.7%) had between 1 and 27 metacercariae of C. complanatum (mean intensity of infection = 9.3 +/- 9.6). In March 1999 and April of both 2000 and 2001, the specimens examined were not infected. The infected fish had a total weight higher than the expected value (Kn > 1.0), while the non-infected fish had a weight equal to the expected value (Kn = 1.0).     

Sequences of 596 cDNA Clones (565,977 bp) of Japanese Flounder (Paralichthys olivaceus) Leukocytes Infected with Hirame Rhabdovirus

We have partially sequenced 785 sequences of 596 independent complementary DNA clones isolated from a cDNA library of Japanese flounder leukocytes infected with hirame rhabdovirus. These sequences consist of a total of 565,977 base pairs. The average size of the sequenced lengths was 721 bp. Of 596 clones, 386 (64.8%) were identified as previously reported genes by the BLASTN and BLASTX programs. About 30% of the identified clones could be recognized by only the BLASTX program. A total of 251 distinct genes were identified, and 181 of these genes are the first such genes reported from the teleostei. Approximately 27% of the identified Japanese flounder genes appear to be associated with cell division, cell structure or motility, and basic energy metabolism, 29% with gene or protein expression, and 17% with cell signaling, cell communication, and cell or organism defense. The most frequently identified expressed sequence tags of leukocytes of Japanese flounder were gelatinase b and ribosomal protein L23, which both had 1.34% prevalence. Received February 12, 1999; accepted April 13, 1999     

Winter survival of immature instars of Mansonia indubitans Dyar & Shannon and Mansonia titillans Walker (Diptera: Culicidae), in Buenos Aires, Argentina

We conducted a whole year research on the ecology of Mansonia indubitans and Ma. titillans in Macáes Pond, Costanera Sur Reserve, Buenos Aires, Argentina. The usage of different floating plants by immature instars and their overwintering was analyzed. The percentage of usage of the available floating macrophytes (Pistia, Limnobium, and Salvinia) by the larvae and pupae was studied. Also, we defined positivity (P+) as the percentage of plants with immature instars for each plant genus on a monthly basis. Ma. immature instars were captured throughout the year and Pistia was the resource most commonly exploited by the mosquitoes. The percentage of fourth-instar larvae and pupae on Pistia roots with respect to total immature instars captured was assessed on a monthly and seasonal basis. The proportion of fourth-instar larvae and pupae from both species of Mansonia on water lettuce roots, showed significant differences between months and seasons. Our results suggest that the populations of Ma. indubitans and Ma. titillans in Macáes Pond, survive during winter mainly as fourth-instar larvae.     

Epidemiologic characteristics of human campylobacteriosis in the County Primorsko-goranska (Croatia), 2003-2007

The aim of the study was to investigate campylobacteriosis incidence in the County Primorsko-goranska (Croatia) between 2003 and 2007 and to find out possible connection with environmental factors (the average monthly temperature and total monthly precipitation). The data (number of stool samples examined, age and sex distribution of patients, monthly distribution of isolates and distribution of isolates according to the species) from the Laboratory for Diagnostics of Enteric Infections of the Teaching Institute of Public Health of the County Primorsko-goranska (Croatia) were analyzed retrospectively. During the observed period 30,164 stool samples were examined for Campylobacter spp. Campylobacters were identified in 1,242 (4.12%) samples. The overall annual campylobacter incidence rate was 81.3 +/- 21.9/100,000 population. Campylobacter jejuni was found in 1,093 (88%) and C. coli in 149 (12%) patients. Our findings showed age distribution of patients typical for developed countries. The patients were mostly children under 5 years (484.4 +/- 129.1/ 100,000, p < 0.001) and between 5 and 9 years of age (226.5 +/- 60.5/100,000, p < 0.05). Male consistently experienced higher rates, but the difference between genders was significant in the age groups from birth till late twentieth (p < 0.001). Campylobacter rates were significantly associated with monthly average temperatures (p < 0.05), but not with precipitation. Further investigations into the incidence of campylobacteriosis on the national level are necessary. The causes of the noticed monthly distribution, sources of infection and connection with the routes of transmission in humans need to be elucidated as well.     

Population dynamics of the philometrid nematode Margolisianum bulbosum in the southern flounder Paralichthys lethostigma (Pisces: Paralichthyidae) in South Carolina, USA

This is the first report of the philometrid nematode Margolisianum bulbosum Blaylock and Overstreet, 1999 from the southern flounder Paralichthys lethostigma on the east coast of the USA. Observation of adult female worms was used as an indication of the parasite's presence in the fish. Adult females were found only in P. lethostigma >50 mm total length. The overall prevalence was 74%, with a mean intensity of 5 female nematodes per parasitized fish. Infected flounders were found throughout the year with a statistically significant decrease in intensity in the winter months. Neither salinity, water temperature, fish gender nor fish age were found to influence either prevalence or intensity of infection in the flounder. While larvigerous (gravid) females were found throughout the year, the significant decrease in their occurrence during the summer through fall, in concert with an observed decrease in intensity of infection during the winter, indicated that the life cycle of this philometrid species is likely to be annual.     

An outbreak of Trichophyton mentagrophytes infection in a rat breeding stock and its successful control

An outbreak of dermatophytosis by Trichophyton mentagrophytes infection was observed in a breeding stock of the Wistar-Imamichi rat. Typical cutaneous lesions such as alopecia and hyperkeratosis due to the fungus were observed in 10% and 44% of adult females, and males respectively. No infection was seen in infant, even in those fostered by infected females. By 3 repeated cultivations of the hair, T. mentagrophytes was isolated from 107 (90.7%) of 118 rats clinically diagnosed as dermatophytosis. In order to maintain the original stock, the usual wholesale slaughter program was avoided and the following control measures were taken. After sacrificing all rats with the cutaneous lesions, the breeding colony was transferred temporarily to a separate animal room. The resulting vacant room was then disinfected 3 times with 10% formalin, followed by spraying with 3% sodium propionate solution 3 month later. The progenies from the non-infected mothers were dipped into 3% sodium propionate solution and transferred back into the disinfected animal room for re-colonization. Frequent sterilization of all equipment with iodophor, balsam and formalin was carried out. The room sterilized monthly with iodophor and 70% alcohol. Daily observation of all rats and monthly screening by culture test was done thereafter. As a result, no infection has been detected 4 month after recolonization, except for the 3 rats which were sacrificed immediately after detection. On the basis of the culture test and clinical observation, no infection was observed in the treated rat colony for the next five following years. Therefore, the effectiveness of these procedures of the eradication of T. mentagrophytes infection from a rat colony was shown to be quite reliable.