CERCARIAL LONGEVITY AND INFECTIVITY OF BOLBOPHORUS DAMNIFICUS, WITH NOTES ON METACERCARIAL PERSISTENCE AND SITE SPECIFICITY IN CHANNEL AND HYBRID CATFISH.

Advances in hybridization practices in U.S. catfish aquaculture have led to increased production of channel (Ictalurus punctatus) × blue catfish (Ictalurus furcatus) hybrids to capitalize on their more favorable production characteristics. However, the effects of typical channel catfish pathogens on hybrids are not well understood, including the digenean Bolbophorus damnificus, which has caused significant losses in channel catfish production. Three experiments were conducted to assess the longevity and site specificity of 2 life stages of B. damnificus impacting catfish production. The first experiment investigated the cercarial longevity and infectivity of B. damnificus over time. Channel catfish were individually challenged with 100 cercariae/fish with cercariae aged in 12-hr time intervals over 5 days (n = 5 fish/time point), with metacercarial cysts excised and enumerated 14 days postchallenge. There was a decrease in cercaria viability and encysted metacercariae over the first 36 hr, with the 12-hr time point having both the greatest cercaria survival and the highest number of metacercariae in exposed fish. The second experiment investigated the longevity of metacercariae within both channel and hybrid catfish. Fish (n = 30) were exposed to 2 treatments (75 or 150 cercariae/fish), and 2 fish from each treatment were sampled every 3 mo for 13 mo. Live metacercariae, based on motility observed after excystment, were found in both species up to 13 mo postchallenge, indicating the metacercariae of B. damnificus can persist throughout an entire growing season in both channel and hybrid catfish. The third experiment investigated the site specificity of metacercariae within both channel and hybrid catfish. Fish (n = 60/species) were challenged with 300 cercariae/fish and 9 fish/species were sampled after 90 days. Metacercariae were excised and enumerated from the anterior midsection (head and body), posterior midsection (trunk/caudal peduncle), ventral (belly), and caudal fin (tail) sections of each fish. Overall, the trunk/caudal peduncle had a 2-fold increase in the number of metacercariae excised, and although not significantly higher, results indicate this region should be the focal point of pondside assessment for the presence of B. damnificus because of ease of detection of encysted metacercariae.

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

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

Abnormal microstructure of the atrophic thalamus in preterm survivors with periventricular leukomalacia.

BACKGROUND AND PURPOSE: The neuroanatomic substrate of cognitive deficits in long-term survivors of prematurity with PVL is poorly understood. The thalamus is critically involved in cognition via extensive interconnections with the cerebral cortex. We hypothesized that the thalamus is atrophic (reduced in volume) in childhood survivors of prematurity with neuroimaging evidence of PVL and that the atrophy is associated with selective microstructural abnormalities within its subdivisions. MATERIALS AND METHODS: We performed quantitative volumetric and DTI measurements of the thalamus in 17 children with neuroimaging evidence of PVL (mean postconceptional age, 5.6 ± 4.0 years) who were born prematurely and compared these with 74 term control children (5.7 ± 3.4 years). RESULTS: The major findings were the following: 1) a significant reduction in the overall volume of the thalamus in patients with PVL compared with controls (P < .0001), which also correlated with the severity of PVL (P = .001); 2) significantly decreased FA (P = .003) and increased λ(⊥) (P = .02) in the thalamus overall and increased axial, radial, and mean diffusivities in the pulvinar (P < .03), suggesting injury to afferent and efferent myelinated axons; and 3) a positive correlation of pulvinar abnormalities with those of the parieto-occipital white matter in periventricular leukomalacia, suggesting that the pulvinar abnormalities reflect secondary effects of damaged interconnections between the pulvinar and parieto-occipital cortices in the cognitive visual network. CONCLUSIONS: There are volumetric and microstructural abnormalities of the thalamus in preterm children with PVL, very likely reflecting neuronal loss and myelinated axonal injury. The selective microstructural damage in the pulvinar very likely contributes to abnormal cognitive visual processing known to occur in such survivors.

Loss of epithelial cell surface carbohydrates during experimental oral carcinogenesis in the rat.

Cell surface glycoconjugates were investigated in a rat model of oral chemical carcinogenesis. The lectins Griffonia simplicifolia (GS-I-B4; specific for alpha-D-galactosyl end groups) and Ulex europeus (UEA-I; specific for alpha-L-fucosyl groups) were examined microspectrofluorimetrically in the oral epithelium of rats painted with the carcinogen 4-nitroquinoline N-oxide (4NQO) and compared with those treated with solvent alone. After labelling with GS-I-B4, the fluorescent intensity of the basal and parabasal epithelial cells was significantly less after 9 months of 4NQO treatment and in overt squamous cell carcinomas compared to controls. The fluorescent activity of the spinous epithelial cells in the non-invasive tissues treated with 4NQO and in the well differentiated (sites of keratin elaboration) malignant epithelium of squamous cell carcinomas was unchanged after labelling with UEA-I. UEA-I failed to stain undifferentiated (areas lacking keratin) malignant epithelium. The findings indicate that alpha-D-galactosyl residues are diminished on the membranes of premalignant and malignant rat epithelial cells. The expression of alpha-L-fucosyl groups, however, remains unchanged in premalignant rat oral epithelium and is closely correlated to the presence of keratin in the malignant epithelium of squamous cell carcinomas.

The expression of cell surface MHC class I heavy and light chain molecules in pre-malignant and malignant lesions of the oral mucosa.

Major histocompatibility complex (MHC) class I antigenic expression was examined on epithelial cell surfaces in normal oral mucosa, non-specific oral keratoses, dysplastic epithelium adjacent to carcinomas and in invasive tumour islands of squamous cell carcinomas, using antibodies to HLA-A,B,C shared determinant antigen and beta 2 microglobulin (beta 2m). HLA-A,B,C antigens were present in the basal and lower spinous cells in normal and dysplastic epithelium and in the non-specific keratoses, but in a minority of carcinomas staining was disorganized and absent at the periphery of tumour islands. beta 2m staining was present in the basal and lower spinous epithelial cells in all tissues; staining was lost progressively with increasing dilution of the primary antibody until it was completely lost in the invasive carcinoma tissue at 1:400, in dysplastic epithelium at 1:800 and in non-specific keratoses at 1:3200; in contrast, staining persisted in normal tissues at greater than 1:3200 anti-beta 2m dilution. Loss of beta 2m staining in the dysplastic epithelial tissues correlated broadly with the degree of cellular atypia. The results suggest that there are decreased concentrations of cell surface beta 2m in potentially malignant and malignant epithelial tissues, with normal concentrations of MHC class I heavy chains.

Oral epithelial atypia and acantholytic dyskeratosis in rats painted with 4-nitroquinoline N-oxide.

Oral epithelial atypia and foci of acantholytic dyskeratosis (FAD) were investigated in 54 rats treated with the carcinogen 0.5% (w/v) 4-nitroquinoline-N-oxide in propylene glycol and in 18 rats treated with propylene glycol only. The palate of each animal was painted 3 times weekly for up to 9 months and rats were killed at monthly intervals. A gradual significant increase in the epithelial atypia indices of the palatal and lingual tissues (anterior and posterior of the intermolar tubercle) was observed with a maximum value of 17-22 of a possible 75 at 28-32 weeks. No significant differences were noted between the atypia indices of the palatal and lingual tissues. FAD were not evident in the palate or lingual tissues before 12 weeks and 16-24 weeks, respectively, and although the palate consistently showed more FAD compared with the lingual tissues the differences were not significant. Features of epithelial atypia and FAD were absent in the 18 control rats treated with propylene glycol only and in 8 untreated control animals. At 28 weeks of 4NQO treatment 2 of 5 rats, at 32 weeks 3 of 4 rats and at 36 weeks 3 of 3 rats had developed infiltrating squamous cell carcinomas in either/both the palate or tongue. The results suggest that epithelial dysplasia and acantholytic dyskeratosis may be late morphological features of a more fundamental change occurring earlier in the process of tumour development.

The use of the lectin Ulex europeus to study epithelial cell differentiation in neoplastic and non-neoplastic oral white lesions.

Ulex europeus (Ulex 1) has been used to study the expression of cell surface alpha-L-fucose on oral epithelial cells of normal human mucosa, white lesions and carcinomas. In normal tissue and in 11 of 12 specimens of non-specific keratoses (leukoplakia), Ulex 1 stained epithelial spinous cells only. In 16 specimens of oral lichen planus, Ulex 1 labelled spinous epithelial cells, and in 12 there was also staining of the basal epithelial cells. Two specimens of white sponge naevus showed a total absence of epithelial staining with Ulex 1. Squamous carcinomas showed a loss of staining in the invading epithelial islands in six of eight specimens, but normal labelling of the surfaces of spinous cells at sites distant from the areas of epithelial invasion. When the severity of epithelial dysplasia, as assessed by the index of epithelial atypia, was compared with the pattern of staining of Ulex 1 in normal mucosa and white lesions there was no correlation. The results show that failure of Ulex 1 to stain cell surface carbohydrates may not be attributable to malignant transformation. The staining of cells not normally stained may be related to hitherto masked residues revealed by cell damage or altered differentiation.