Gokmen Zafer Pekmezci

Molecular identification of Anisakis species (Nematoda: Anisakidae) from marine fishes collected in Turkish waters.

Anisakid nematodes are important etiological agents for zoonotic human anisakiasis (or anisakidosis). These parasites in the Turkish waters still remain unexplored. This study aims the molecular identification of Anisakis species in Turkeys coast from Black, Aegean and Mediterranean Sea and specifically to screen for zoonotic species in commonly commercialized a total of 1145 fish belonging to 31 different species using both polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) and sequencing of the ribosomal internal transcribed spacer (ITS) regions and the mitochondrial cytochrome C oxidase subunit II (cox2) gene. A total of 776 Anisakis type I larvae were isolated in 56/1145 (4.8%) fish of 7 species from Turkish waters. The combining all of our results, e.g., morphology, PCR-RFLP, ITS region, and the cox2 gene, conclusively supported the identification of 3 Anisakis spp. taken from marine fish hosts, namely Anisakis pegreffii, Anisakis typica and Anisakis simplex sensu stricto (s.str.)/A. pegreffii hybrid genotype. No Anisakis larvae were isolated from the Black Sea whereas A. pegreffii, A. typica and A. simplex s.str./A. pegreffii hybrid genotype was found in the Aegean Sea and A. pegreffii was only isolated from the Mediterranean Sea. This study represents the first identification of A. typica and A. simplex s.str./A. pegreffii hybrid genotypes from Turkish waters. Moreover, in the present study first record of the presence of A. pegreffii is also reported from Turkish coasts of Aegean and Mediterranean Sea. No zoonotic Anisakis species were found in commonly commercialized 1025 fish belonging to 16 different species from the Black Sea, thus Turkish populations who consume captured fish from the Black Sea may have a less risk of human anisakiasis or allergies. However, the prevalence of larvae were 47.1% and 46% and recognized zoonotic A. pegreffii were identified from the Aegean and Mediterranean Sea coast, suggesting a high threat of anisakiasis or allergies for Turkish populations who consume fish originating in these regions.

Molecular characterization of Hysterothylacium fabri (Nematoda: Anisakidae) from Zeus faber (Pisces: Zeidae) caught off the Mediterranean coasts of Turkey based on nuclear ribosomal and mitochondrial DNA sequences.

In the present study, Hysterothylacium fabri was found in the coasts of the Mediterranean Sea, Turkey and characterized by sequencing of nuclear (internal transcribed spacer, ITS) and mitochondrial (cytochrome c oxidase subunit 2, cox2) markers. Pairwise comparison between the entire ITS fragment including ITS-1, 5.8S, ITS-2 sequences of the H. fabri isolates from the Mediterranean Sea (Turkey, KC852206) and other H. fabri isolates from the South China Sea (JQ520158), the South Korea waters (JX974558) showed differences ranged from 0.1 and 1.1%. With the present study, H. fabri from the Mediterranean Sea was characterized for the first time by sequencing of the cox2 gene.

Occurrence of Anisakis simplex sensu stricto in imported Atlantic mackerel (Scomber scombrus) represents a risk for Turkish consumers

Anisakid larvae are a prevalent food-borne pathogen that has been found in numerous fish species destined for human consumption. The accidental consumption of infected raw or poorly cooked fish may cause gastroenteric diseases and allergies in humans. In spite of the fact that thorough cooking or freezing kills Anisakis worms, this method does not destroy their allergenic capacity. The presence of A. simplex (s.s.) in seafood products may present a health risk for consumers. In Turkey, Atlantic mackerels are marketed as frozen and mainly imported from Norway. The aim of this study was to identify the Anisakis species found in deep-frozen whole Atlantic mackerel (Scomber scombrus) destined for human consumption in fish markets that imported fish from Norway to Turkey. All Anisakis larvae isolated from imported Atlantic mackerel were identified via morphology as third larvae of Anisakis Type I. The ITS region (ITS-1, 5.8S subunit, ITS-2) was amplified and digested with the restriction enzymes Hinf I and Hha I. Larvae of the genus Anisakis were identified via PCR-RFLP as belonging to Anisakis simplex (s.s.), and this was confirmed by sequencing the cox2 gene. The overall prevalence of Anisakis larvae was 25% (95% confidence limits: 13-41%), and the mean intensity was 19.1 (bootstrap 95% confidence limits: 15.3-25.5). Recognized zoonotic A. simplex (s.s.) larvae found in imported Atlantic mackerel could represent a risk. Those who consume them could acquire parasitic allergies. The results will have an important impact on public health risk assessment in that they suggest reviewing critical control points at the Hazard Analysis Critical Control Point (HACCP) programmer to reduce the risk of anisakid-induced allergies among consumers. Consequently, the present study provides the first data regarding the occurrence of A. simplex (s.s.) larvae in imported Atlantic mackerel in Turkish markets.

Myxobolus anatolicus sp. nov. (Myxozoa) infecting the gill of Anatolian khramulya Capoeta tinca (Cyprinidae) in Turkey.

This work is part of an ongoing investigation into the characteristics of myxozoan parasites of freshwater fish in Turkey and was carried out using morphology, histopathology and molecular analysis. A new species of the genus Myxobolus (M. anatolicus sp. nov.) was found infecting the gills of 3 of 34 specimens (8.8%) of Anatolian khramulya Capoeta tinca from the Samsun Province, Northern Turkey. Both morphology and 18S rDNA sequence data revealed that M. anatolicus sp. nov. was distinct from other Myxobolus species found in the gills of cyprinid fishes. The small, white and round-shaped plasmodia, measuring 0.2 to 1.4 mm in diameter, were observed macroscopically in the gills. Histological analysis revealed that the cyst-like plasmodia have an intralamellar-vascular type development. Mature spores of M. anatolicus sp. nov. were oval in both frontal and sutural views, and tapered at the anterior poles. The spores were 10.1 ± 0.41 (9.4 to 10.7) µm long, 6.9 ± 0.28 (6.6 to 7.2) µm wide, and 4.5 ± 0.36 (4.4 to 4.6) µm thick. The 2 polar capsules were pyriform, equal in size, 4.6 ± 0.45 (4.4 to 4.8) µm long and 2.1 ± 0.12 (2 to 2.3) µm wide. Polar filaments within the polar capsules were coiled with 5 or 6 turns. Phylogenetic analysis placed M. anatolicus sp. nov. in a clade of gill-infecting myxobolids. This is the first record of a Myxobolus species infecting Anatolian khramulya Capoeta tinca, and the first record of this species from Eurasia.

Immunohistochemical evaluation of experimental Vagococcus salmoninarum infection in rainbow trout (Oncorhynchus mykiss, Walbaum 1792)

The aim of this study was to investigate the pathogenesis and histopathological and immunohistochemical findings in rainbow trout (Oncorhynchus mykiss) following experimental vagococcosis. For this purpose, 60 rainbow trout were used. The experimental study used the pathogen Vagococcus salmoninarum. The fish were intraperitoneally (IP) administered with an inoculate containing 0.1 mL of the bacteria, resulting in a dose of 1.2 × 10(9) cfu mL(-1) per fish. For histopathological observations, tissue samples were taken from fish that died during the experiment and fish that survived until the end of the trial (60th day). All the tissue samples were immunohistochemically stained by the avidin-biotin-peroxidase complex and immunofluorescence methods using polyclonal antibody to detect V. salmoninarum antigens. In immunoperoxidase staining, positive reactions to bacterial antigens were most commonly seen in the kidney, heart and liver. In the immunofluorescence analysis, the distribution of antigens in the tissue and organs was similar to that observed with the immunoperoxidase staining. The results reveal an important correlation between histochemical and immunohistochemical staining in demonstrating the distribution of V. salmoninarum antigens in the affected tissues.

Efficacy of amitraz plus inactivated parapoxvirus ovis in the treatment of canine generalised demodicosis

Canine generalised demodicosis (CGD) is a challenging disease to treat effectively. Inactivated parapoxvirus ovis (iPPVO) could help to accelerate treatment with acaricidial therapy by altering the immune response. This study was designed to investigate the effects of treating CGD with amitraz plus iPPVO in terms of clinical outcomes and blood parameters. The study involved 16 dogs ranging in age from eight months to six years and weighing between 10 and 40 kg. Eight dogs were treated with amitraz and eight with amitraz plus iPPVO. Biochemical analysis of whole blood and serum, including serum C reactive protein (CRP) and serum amyloid A (SAA), was performed. Skin scrapings were conducted on days 0, 10, 40, 80 and 120 of treatment, and mite numbers were recorded. Clinical remission was determined according to mite numbers and clinical scores. The difference in mean whole remission days between the amitraz group (104.3 days) and the amitraz+iPPVO group (84.5 days) was statistically significant (P<0.05). Mean clinical scores were also significantly better in the amitraz+iPPVO (5.60) group when compared with the amitraz group (7.65). No adverse reactions were observed in either group. In view of these findings, the use of iPPVO in conjunction with amitraz can be recommended for treating CGD.

Occurrence and molecular characterization of Clinostomum complanatum (Trematoda: Clinostomidae) in freshwater fishes caught from Turkey

The metacercariae of Clinostomum species which known as yellow grubs have zoonotic potential by infecting humans. In the present study, a total of 403 freshwater fish specimens belonging to different genera from Central Anatolia Region of Turkey were examined for yellow grub metacercariae infections. Only three specimens belonging to Squalius cephalus were found to be infected with metacercariae with a prevalence on this host species of 2.4% and an overall prevalence of 0.7%. All the metacercariae were morphologically identified as Clinostomum complanatum. Partial fragments of mitochondrial cytochrome oxidase I (mt-COI) gene and internal transcribed spacer 2 (ITS-2) were amplified for sequence and phylogenetic analyses. The sequence analyses of ITS-2 and mt-COI revealed three and nine polymorphic sites leading to detection of four and five haplotypes within the related gene regions, respectively. Moreover, the intraspecific genetic distances for C. complanatum isolates ranged from 0.0 to 0.7% for ITS-2 and 0.0 to 1.4% for mt-COI data sets. Consequently, the present study has provided first combined morphologic and molecular data on C. complanatum infecting Turkish freshwater fishes.

Ichthyobodo spp. Infection in Meagre (Argyrosomus regius) from Turkey: Parasitological and Pathological Findings.

OBJECTIVE The aim of this study is to describe the first report of Ichthyobodo spp. infection in meagre (Argyrosomus regius) fry in a marine aquaculture facility in Turkey. METHODS The material of the study was composed of 30 meagre A. regius in 2-3 g weight taken from the fry adaptation unit of a fish farm in the Aegean Sea. In this study, parasitological and pathological examinations were performed on the meagre. Ichthyobodo spp. was determined on the body surfaces and gills. RESULTS Pathological examination revealed grayish mucous and erosions between the pin head and lentin over the skin of the examined specimens. Microscopic examinations revealed significant spongiosis, vacuolar degeneration, and hyperplasia in epidermal malpighian cells and hyperplasia in goblet cells. CONCLUSION In the present study, Ichthyobodo spp. infection was for the first time determined in an alternative cultured meagre in Turkey.

Serum concentration and skin tissue expression of insulin-like growth factor 2 in canine generalized demodicosis.

BACKGROUND There is increasing evidence that insulin-like growth factor-2 (IGF-2) levels are altered in skin injury; there are no data evaluating the serum concentration and skin tissue expression of IGF-2 in canine generalized demodicosis. HYPOTHESIS/OBJECTIVES To assess serum concentrations of IGF-2 collected from dogs with generalized demodicosis compared to healthy dogs and to determine the location of IGF-2 in the skin of affected dogs. METHODS Blood and skin samples were collected from 12 dogs of differing breeds and gender at 1-2 years of age that had a confirmed diagnosis of generalized demodicosis. Age-matched control skin and blood samples were collected from 11 normal dogs of different breeds and gender. Serum IGF-2 concentrations were measured by enzyme-linked immunosorbent assay. Skin tissue expression of IGF-2 was analysed by immunohistochemical methods. RESULTS Serum concentration and skin tissue expression of IGF-2 were increased in dogs with generalized demodicosis compared with control dogs. CONCLUSIONS AND CLINICAL IMPORTANCE These findings indicate that keratinocytes, histiocytes and fibrocytes in the dermis are positive for IGF-2; they may be a source of the elevated serum IGF-2 levels in dogs with generalized demodicosis.

A case of auricular, anal and umbilical myiasis caused by the larvae of Phormia regina (Meigen) (Diptera: Calliphoridae) in neonatal kittens.

The occurrence of feline myiasis is rare. Massive infestations of dipteran larvae can lead to death if not treated at an early stage. Auricular, anal and umbilical myiasis was detected in three neonatal kittens. The dipteran larvae were collected, fixed in 70% alcohol and clarified with 10% KOH for a few days. Later, larvae were dissected under the stereomicroscope, mounted on slides and then identified as the third instar of the black blowfly, Phormia regina (Meigen) (Diptera: Calliphoridae), according to their stigmatic and cephaloskeleton structures. Original measurements and figures are presented. Treatment included mechanical removal of larvae and cleansing of the area by applying polyvinylpyrrolidone-iodine complex. The presence of P. regina in cats has been reported here for the first time in Turkey.