Under the radar: co-introduced monogeneans (Polyopisthocotylea: Gastrocotylinea) of the invasive fish Scomberomorus commerson in the Mediterranean Sea.

The Levant Basin is in many ways the world's most invaded marine ecosystem owing to the existence of the man-made Suez Canal. The invasion of free-living organisms through this pathway is increasingly documented and monitored in the past two decades, and their ecological impact recognized. Nonetheless, while tremendous scientific effort is invested in documenting introduced fishes, co-introduction events of these fishes and their parasites have drawn relatively little interest. In our research, we examined the presence of gill parasites (Monogenea) on the invasive narrow barred Spanish mackerel Scomberomorus commerson which has been known in the Mediterranean Sea for 80 years. The gills of S. commerson supported numerous, relatively large monogeneans (Monogenea: Gastrocotylinea), reaching prevalence levels of 100% with a mean intensity of ~ 80 worms per host. Using an integrated molecular and morphological approach, four gastrocotylinean species were identified: Gotocotyla acanthura, Cathucotyle cathuaui, Pricea multae, and Pseudothoracocotyla ovalis. Two species, C. cathuaui and P. ovalis, are reported here for the first time from the Mediterranean. Sequences of the 28S rRNA gene of G. acanthura from native hosts, Pomatomus saltatrix and Trachinotus ovatus, differed from individuals collected from S. commerson by 1.8%. We therefore suggest that the taxonomic status and distribution of G. acanthura should be revisited, and we recommend an integrated approach as essential to accurately detect co-introductions.

Distribution and population structure of the alien Indo-Pacific Randall's threadfin bream Nemipterus randalli in the eastern Mediterranean Sea.

As part of a comprehensive study of trawl fishery catch off Israel (Ashdod) and Turkey (Iskenderun and Antalya) conducted during 2008-2011, the population explosion of Nemipterus randalli, first recorded in the Mediterranean Sea in the beginning of 2005, was documented. The smallest individuals occurred on deeper bottoms (120 m), significantly more individuals were collected at night, and juvenile recruitment to the commercial fishery occurred during November and December at 40 m depth.

Characterization of swim bladder non-inflation (SBN) in angelfish, Pterophyllum scalare (Schultz), and the effect of exposure to methylene blue.

Failure to inflate the swim bladder is regarded a major obstacle in the rearing of many fish species. We present a study of swim bladder non-inflation (SBN) in angelfish, Pterophyllum scalare. A normal developing primordial swim bladder was first discernable at the end of the first day post-hatch (p.h.) as a cluster of epithelial cells with a central lumen, surrounded by presumably mesenchymal cells. Initial inflation occurred on the fourth day p.h. Prior to inflation the swim bladder epithelium consisted of an outer squamous and inner columnar layer. Cells of the inner layer were filled at their basal region with an amorphous material, which disappeared upon inflation. A pneumatic duct was absent, and larvae presented no need to reach the water surface for inflation, suggesting that angelfish are pure physoclists. A model for the role of the amorphous material in normal initial inflation is proposed. Abnormal swim bladders were apparent from the fourth day p.h., and methylene blue (MB) at a concentration of 5 ppm significantly increased the prevalence of SBN. Histologically, abnormal swim bladders in larvae hatched in 5 ppm MB could not be distinguished from those in fish raised under routine conditions (0.5 ppm MB). We suggest that MB may have a teratogenic effect in angelfish.

An unidentified epi-epithelial myxosporean in the intestine of gilthead sea bream Sparus aurata L.

In the course of experimental infections of gilthead sea bream Sparus aurata with the myxozoan Enteromyxum leei, stages of an unidentified myxozoan were observed attached to the intestinal brush border of some fish. Infection levels of the parasite, which was named "epi-epithelial myxosporean" (EEM) were recorded, and its structure was studied by light microscopy (LM) and electron microscopy (EM). In situ hybridisation (ISH) probes specific for E. leei were developed and used to differentiate between the two parasites. The EEM parasite was observed only in epi-epithelial position on the intestine mucosa and never in any of the other tissues studied (kidney and gall bladder). Prevalence was variable, with values reaching 40.2%. With transmission EM, trophozoites displayed pseudopodia-like projections inserted in between the enterocyte microvilli, producing an intimate interface. No mucosal histopathology that could be attributed to the myxozoan was found. EEM stages did not stain with the E. leei-specific ISH probes. From the results of the LM, EM and ISH studies, we conclude that the EEM parasite found in gilthead sea bream intestine in both Mediterranean and Red Sea sites is a coelozoic myxosporean, distinct from E. leei.

Experimental transmission of Enteromyxum leei to freshwater fish.

The myxosporean Enteromyxum leei is known to infect a wide range of marine fish hosts. The objective of the present study was to determine whether freshwater fish species are also receptive hosts to this parasite. Seventeen species of freshwater fish were experimentally fed E. leei-infected gut tissue from donor gilthead sea bream Sparus aurata obtained from a commercial sea bream cage farm. Four of the tested species, tiger barb Puntius tetrazona, zebra danio Danio rerio, oscar Astronotus ocellatus and Mozambique tilapia Oreochromis mossambicus, were found to be susceptible with prevalences ranging from 53 to 90%. The course of infection and pathology was limited to the gut mucosa epithelium and was similar to that observed in marine hosts. Little is known of the differences in physiological conditions encountered by a parasite in the alimentary tract of freshwater vs. marine teleost hosts, but we assume that a similar osmotic environment is maintained in both. Parasite infectivity may be influenced by differences in the presence or absence of a true stomach, acidic gastric pH and digestive enzyme activity both in the stomach and intestine. Variability in susceptibility among species may also stem from differences in innate immunity. Dimensions of spores produced in the donor sea bream and recipient freshwater species are variable in size, as previously observed in other captive marine host species.

Hyperparasitism of trichodinid ciliates on monogenean gill flukes of two marine fish.

Two unusual cases of hyperparasitism of trichodinid ciliates on monogenean gill flukes are described from southern Israel (Red Sea). The first case occurred in cultured European sea bass Dicentrarchus labrax infected by Diplectanum aequans, while the second was observed in a feral devil firefish Pterois miles infected by Haliotrema sp. In both cases, the trichodinids heavily co-infested the host fish gills. The flukes were completely coated by the ciliates, which gave them a cobblestone appearance, but no damage to their tegument was apparent. Both cases are most likely a result of accidental hyperparasitism, brought about by perturbed environmental conditions.

Kudoa iwatai (Myxosporea: Multivalvulida) in wild and cultured fish in the Red Sea: redescription and molecular phylogeny.

Gilt-head sea bream, Sparus aurata L., the Mediterranean's most important mariculture species, has been cultured for the last 30 yr in Eilat (Israeli Red Sea). Kudoa sp. was the first myxosporean parasite reported from this species. In recent years, an increase in prevalence in both land-based and sea-cage facilities in Eilat has been observed. Infections with the same Kudoa species appeared in cultured European sea bass Dicentrarchus labrax (L.) and grey mullet Mugil cephalus in the same farms, as well as in 10 species of wild Red Sea reef fish, indicating that Kudoa sp. is not fastidious with regard to its host. All affected species displayed 1- to 2-mm (up to 5 mm) whitish, spherical, or oval polysporous plasmodia. The parasite established multiple site infections, most commonly in the muscles and intracranial adipose tissue of the brain and eye periphery. Other sites were subcutaneous adipose tissue, nerve axons, mouth, eye, mesenteries, peritoneum, swim bladder, intestinal musculature, heart, pericardium, kidney, and ovary. On the basis of spore morphology, the parasite was identified as Kudoa iwatai Egusa and Shiomitsu, 1983. Ultrastructural features were comparable to those of previously studied Kudoa species. The 18S rDNA from 7 Red Sea isolates was sequenced and compared with the sequence of the same gene from K. iwatai isolated from cultured red sea bream, Pagrus major, in Japan. The phylogenetic position of K. iwatai within the genus was determined using sequence analysis of all related taxa available in GenBank. The 3 isolates of K. iwatai clustered together on a newly formed, highly supported clade. The Red Sea strain of K. iwatai is apparently native to the region. In the absence of records of this Kudoa sp. from the extensive Mediterranean sea bream and sea bass production industries, introduction with its Mediterranean hosts seems unlikely. Therefore, we conclude that K. iwatai is an Indo-Pacific species that, in the Red Sea, has extended its host range to include the allochthonous gilt-head sea bream, European sea bass, and grey mullet.

Nodavirus infections in Israeli mariculture.

Viral encephalopathy and retinopathy (VER) infections were diagnosed in five fish species: Epinephelus aeneus, Dicentrarchus labrax, Sciaenops ocellatus, Lates calcarifer and Mugil cephalus cultured on both the Red Sea and Mediterranean coasts of Israel during 1998-2002. Spongiform vacuolation of nervous tissue was observed in histological sections of all examined species. With transmission electron microscopy, paracrystalline arrays and pieces of membrane-associated non-enveloped virions measuring approximately 30 nm in diameter were observed in the brain and retina of all species. At the molecular level, the nodavirus was detected by using a primer set that amplified the T4 region of the coat protein gene. When the same set of primers was used to search for VER in an additional fish species, Sparus aurata, it was found to produce non-specific amplicons, giving rise to false-positive results. This problem was overcome by using a different primer set (F1/VR3), designed on a highly conserved region of the virus gene, which amplified a fragment of 254 bp, and confirmed that S. aurata was nodavirus-free. This set was validated on all five species of infected fish, as well as clinically healthy fish. Comparison of the coat protein genes from the Israeli isolated sequences indicated that more than one viral strain was involved. No strict host-specificity was evident. Red Sea and Mediterranean isolated sequences grouped in distinct clusters, together with several foreign isolates from the Mediterranean area and the Far East, as phylogenetically close to the Epinephelus akaara RGNNV type.

A new species of Sphaeromyxa (Myxosporea: Sphaeromyxina: Sphaeromyxidae) in devil firefish, Pterois miles (Scorpaenidae), from the northern Red Sea: morphology, ultrastructure, and phylogeny.

Sphaeromyxa zaharoni n. sp. (Myxosporea) is described from the gallbladder of devil firefish, Pterois miles (Scorpaenidae), from coral reefs of the Gulf of Eilat, Israel, northern Red Sea. The parasite was found also in bearded scorpionfish, Scorpaenopsis barbata, from the same area. This is the first report on Sphaeromyxa sp. from this zoogeographical region. The plasmodia are amoeboid when young, becoming disc-shaped and elongated when mature. In paraffin sections, the plasmodium periphery appears as a finely granulated, strongly eosinophilic layer with an intricate surface membrane network. Sphaeromyxa zaharoni n. sp. is polysporous, disporoblastic, and has asynchronous sporogenesis. The mature spore is elongated and fusiform, has delicately ridged valves, and contains a single, binucleated sporoplasm. In valvular view, the tips are truncated. The mean spore size is length 14.5 microm, width 4.8 microm, and polar capsule 4.8 x 3.4 microm. The 2 equally sized ovoid polar capsules are positioned at opposite ends of the spore, each containing a filament loosely folded in 2 loops. The fine structure of the sporoblast and spore corresponded with previously studied Sphaeromyxa species. According to small-subunit ribosomal DNA gene sequence analysis, S. zaharoni n. sp. is most closely related to 2 Myxidium spp. The close phylogenetic relatedness of Sphaeromyxa and Myxidium and similar spore morphology raises the question whether these 2 genera should be maintained in separate families and suborders.

Lactococcus garvieae in wild Red Sea wrasse Coris aygula (Labridae).

Lactococcus garvieae infection in wild wrasse Coris aygula is reported, and the serological and molecular characteristics of the isolate are described. This is the first evidence of the presence of this pathogen in the Red Sea, and it follows the recent diagnosis of Mycobacterium marinum and Streptococcus iniae in wild fish from the same region. Whether all 3 pathogens are strains endemic to the Red Sea, or recent introductions into the region, remains to be determined, but their appearance over a period of a few years in wild fish populations in the northern Red Sea is consistent with an emerging trend affecting marine organisms on a global level in areas subjected to intense anthropogenic impacts.

Trematode metacercariae of fishes as sentinels for a changing limnological environment.

Trematode metacercaria populations infecting cichlids in Lake Kinneret (Sea of Galilee) were used as sentinels for the changing limnological environment. Parasitological data from 0+ cichlid fingerlings (Tilapia zillii, Oreochromis aureus, Sarotherodon galilaeus) were collected from the northwest shore of the lake during 1999 to 2001 and compared with data obtained from 1982 to 1984. The results show that the composition of the metacercarial populations changed significantly between the 2 samplings periods. The total number of metacercarial species found in the Kinneret cichlids was lower in 1999 to 2001 than in 1982 to 1984. Metacercariae transmitted by the snail Bulinus truncatus (Clinostomum tilapiae, Euclinostomum heterostomum, Bolbophorus levantinus and Neascus-type metacercariae--black spot and others) that were commonly found in 1982 to 1984 were absent from the fishes sampled during 1999 to 2001. The other trematode metacercariae (Centrocestus sp. and Haplorchis sp. transmitted by Melanoides tuberculata, Pygidiopsis genata transmitted by Melanopsis costata, as well as Phagicola longa, Strigeidae sp.1, an unidentified metacercaria typically found in the liver, and glochidia) increased in abundance in fishes sampled during 1999 to 2001. The calculated 'true' species richness of the habitat, extrapolated as a function of sampling effort, was significantly lower in the 1999 to 2001 versus 1982 to 1984 samples, while significantly fewer fishes needed to be sampled ('sampling effort') during 1999 to 2001 in order to reach the 'true' species richness. The higher calculated values of species richness and diversity for the 1999 to 2001 samples despite the overall decline in species number is explained by the overall increase in metacercariae prevalence.

Strain variation in Mycobacterium marinum fish isolates.

A molecular characterization of two Mycobacterium marinum genes, 16S rRNA and hsp65, was carried out with a total of 21 isolates from various species of fish from both marine and freshwater environments of Israel, Europe, and the Far East. The nucleotide sequences of both genes revealed that all M. marinum isolates from fish in Israel belonged to two different strains, one infecting marine (cultured and wild) fish and the other infecting freshwater (cultured) fish. A restriction enzyme map based on the nucleotide sequences of both genes confirmed the divergence of the Israeli marine isolates from the freshwater isolates and differentiated the Israeli isolates from the foreign isolates, with the exception of one of three Greek isolates from marine fish which was identical to the Israeli marine isolates. The second isolate from Greece exhibited a single base alteration in the 16S rRNA sequence, whereas the third isolate was most likely a new Mycobacterium species. Isolates from Denmark and Thailand shared high sequence homology to complete identity with reference strain ATCC 927. Combined analysis of the two gene sequences increased the detection of intraspecific variations and was thus of importance in studying the taxonomy and epidemiology of this aquatic pathogen. Whether the Israeli M. marinum strain infecting marine fish is endemic to the Red Sea and found extremely susceptible hosts in the exotic species imported for aquaculture or rather was accidentally introduced with occasional imports of fingerlings from the Mediterranean Sea could not be determined.

Streptococcus iniae infections in Red Sea cage-cultured and wild fishes.

Streptococcus iniae was isolated from 2 moribund wild Red Sea fishes, Pomadasys stridens (Pomadasyidae) and Synodus variegatus (Synodontidae), both collected in shallow waters along the Israeli coast of the Gulf of Eilat. The site is approximately 2 km from a mariculture cage farm in which streptococcal infections were diagnosed in previous years in the red drum Sciaenops ocellatus. This is the first report of S. iniae in Red Sea fishes. Biochemical and molecular similarities between the isolates from cultured fishes and those from the wild specimens suggest that a single strain is involved, and that 'amplification' and dispersal of this pathogen from captive to feral fishes have occurred. At the molecular level, the pathogen is different from the S. iniae isolates that have been afflicting the Israeli freshwater aquaculture in recent years. Although S. iniae prevalence in the wild fish populations of the area remains to be determined, the northernmost region of the Gulf of Eilat, virtually landlocked and with generally calm seas and weak currents, seems to be particularly vulnerable to the impact of diseases that develop in this mariculture system.

Ultrastructure and small-subunit ribosomal DNA sequence of Henneguya lesteri n. sp. (Myxosporea), a parasite of sand whiting Sillago analis (Sillaginidae) from the coast of Queensland, Australia.

Henneguya lesteri n. sp. (Myxosporea) is described from sand whiting, Sillago analis, from the southern Queensland coast of Australia. H. lesteri displays a preference for the pseudobranchs and is typically positioned along the afferent blood vessels, displacing the adjoining lamellae and disrupting their normal array. The plasmodia appeared as whitish-hyaline, elliptical cysts (mean dimensions 230 x 410 microm) attached to the oral mucosa lining of the hyoid arch on the inner surface of the operculum. Infections of the gills were also found, in which the plasmodia were spherical, averaged 240 x 240 microm in size and were located on the inner hemibranch margin. The parasites lodged in the gill filament crypts and generated a mild hyperplastic response of the branchial epithelium. In histological sections, the plasmodium wall and adjoining ectoplasm appeared as a finely granulated, weakly eosinophilic layer. Ultrastructurally, this section of the host-parasite interface contained an intricate complex of pinocytotic channels. H. lesteri is polysporic, disporoblastic and pansporoblast forming. Sporogenesis is asynchronous, with the earliest developmental stages aligned predominantly along the plasmodium periphery, and maturing sporoblasts and spores toward the center. Ultrastructural details of sporoblast and spore development are in agreement with previously described myxosporeans. The mature spore is drop-shaped, length (mean) 9.1 microm, width 4.7 microm, thickness 2.5 pm, and comprises 2 polar capsules positioned closely together, a binucleated sporoplasm and a caudal process of 12.6 microm. The polar capsules are elongated, 3.2 x 1.6 microm, with 4 turns of the polar filament. Mean length of the everted filament is 23.2 pm. Few studies have analyzed the 18S gene of marine Myxosporea. In fact, H. lesteri is the first marine species of Henneguya to be characterized at the molecular level: we determined 1966 bp of the small-subunit (18S) rDNA. The results indicated that differences between this and the hitherto studied freshwater Henneguya species are greater than differences among the freshwater Henneguya species.

Assessing use of primary health care services by very low-income adults in a managed care program.

OBJECTIVE: To assess the effect of providing free health care services to low-income adults. METHODS: We measured access to primary care services by enrollees with 4 chronic medical conditions in the General Relief Health Care Program (GRHCP), a program designed for adults receiving General Relief (GR). Implemented by the Los Angeles County Health Department in October 1995, the GRHCP is composed of private and public health care facilities. As adults registered for GR, they were asked to complete a baseline health survey, were enrolled in the GRHCP, and assigned a health care provider. A total of 8520 surveys were completed between September and November 1996 (98% response rate). The analyses of this article are limited to individuals (N = 2164) who reported a history of hypertension, diabetes mellitus, a nonresolving cough, or substance dependence. We reviewed medical records to determine whether new GR recipients had visited their designated GRHCP provider within 4 months of enrollment and used multivariate logistic regression to assess the effect of individual patient factors on the use of free health care. RESULTS: A total of 17% of individuals visited their assigned GRHCP provider within 4 months of enrollment. In multivariate analysis, patients were more likely to have made a visit if they were younger than 50 years, were female, were Asian/Pacific Islander, reported needing to see a physician, or had seen a physician within 12 months. CONCLUSIONS: It is not sufficient to merely supply the name and address of a health care provider to this population. More aggressive efforts should be attempted to increase utilization of services for patients with medical conditions responsive to ambulatory care.

Health behaviors, health status, and access to and use of health care: a population-based study of lesbian, bisexual, and heterosexual women.

BACKGROUND: There is a dearth of validated information about lesbian and bisexual women's health. To better understand some of these issues, we used population-based data to assess variations in health behaviors, health status, and access to and use of health care based on sexual orientation. METHODS: Our study population was drawn from a population-based sample of women, the 1997 Los Angeles County Health Survey. Participants reported their sexual orientation and these analyses included 4697 women: 4610 heterosexual women, 51 lesbians, and 36 bisexual women. We calculated adjusted relative risks to assess the effect of sexual orientation on important health issues. RESULTS: Lesbians and bisexual women were more likely than heterosexual women to use tobacco products and to report any alcohol consumption, but only lesbians were significantly more likely than heterosexual women to drink heavily. Lesbians and bisexual women were less likely than heterosexual women to have health insurance, more likely to have been uninsured for health care during the preceding year, and more likely to have had difficulty obtaining needed medical care. During the preceding 2 years, lesbians, but not bisexual women, were less likely than heterosexual women to have had a Papanicolaou test and a clinical breast examination. CONCLUSIONS: In this first population-based study of lesbian and bisexual women's health, we found that lesbians and bisexual women were more likely than heterosexual women to have poor health behaviors and worse access to health care. These findings support our hypothesis that sexual orientation has an independent effect on health behaviors and receipt of care, and indicate the need for the increased systematic study of the relationship between sexual orientation and various aspects of health and health care. Arch Fam Med. 2000;9:1043-1051

Receipt of preventive health care services by lesbians.

BACKGROUND: We measured receipt of age-appropriate preventive health services by lesbians and assessed whether provider and individual characteristics, including disclosure of sexual orientation, are independently associated with receipt of these services. METHODS: A questionnaire was printed in a national biweekly gay, lesbian, and bisexual news magazine, and self-identified lesbians living in all U. S. states (N =6935) responded to the survey. Main outcome variables were receipt of a Pap smear within the preceding 1 and 2 years and, for women aged > or= 50, receipt of a mammogram within the past 1 and 2 years. RESULTS: Fifty-four percent had Pap smears within 1 year and 71% within 2 years, with increasing rates among older and more educated respondents. Seventy percent of respondents aged > or = 50 had a mammogram in the past year, and 83% within 2 years; rates did not vary significantly controlling for education. Sixty percent had disclosed their sexual orientation to their regular health care provider. Controlling for patient and provider characteristics, disclosure was independently associated with receipt of Pap smears, but not mammograms. CONCLUSIONS: It is important for providers to identify their lesbian patients' unmet needs for preventive health care. Additionally, it is important for providers to provide complete and appropriate preventive health care for their lesbian patients. Further research is needed to determine why lesbians are not receiving Pap smears at the recommended rate and whether this disparity is reflective of aspects of cervical cancer screening or indicates a more general problem with access to health care including receipt of preventive services.

Mycobacteriosis in wild rabbitfish Siganus rivulatus associated with cage farming in the Gulf of Eilat, Red Sea.

Infection patterns of Mycobacterium marinum were studied over a period of 3 yr in wild rabbitfish Siganus nivulatus populations associated with commercial mariculture cages and inhabiting various sites along the Israeli Red Sea coastline. Mycobacteriosis was first recorded from the Red Sea in 1990 in farmed sea bass Dicentrarchus labrax and is absent from records of studies on parasites and diseases of wild rabbitfish carried out in the 1970s and 1980s. A sharp increase in the prevalence of the disease in cultured and wild fish in the region has occurred since. A total of 1142 rabbitfish were examined over a 3 yr period from inside mariculture net cages, from the cage surroundings and from several sites along the coast. Histological sections of spleens were examined for presence of granulomatous lesions. Overall prevalence levels of 50% were recorded in the rabbitfish sampled inside the net cages and 39% at the cages' close surroundings, 21% at a sandy beach site 1.2 km westwards, 35% at Eilat harbour 3 km to the south and 42% at a coral reef site about 10 km south of the cages. In addition, 147 fish belonging to 18 native Red Sea species were sampled from 2 sites, the net cage farm perimeter and the coral reef area, and examined for similar lesions. None of those from the coral reef were infected with Mycobacterium; however, 9 of 14 species collected from the cage surroundings were infected. An increase in prevalence of mycobacteriosis in the mariculture farm area was noted from 1995 to 1997. At the same time, a significant increase in prevalence was also apparent at the coral reef sampling site. Two M. marinum isolates from rabbitfish captured at Eilat harbour and the coral reef site were shown by 16S rDNA sequencing analysis to be identical to isolates from rabbitfish trapped inside the mariculture cages as well as isolates from locally cultured sea bass D. labrax. The implications of spreading of M. marinum infection in wild fish populations in the Gulf of Eilat are discussed.