Biofilm and Sediment are Major Reservoirs of Virulent Aeromonas hydrophila (vAh) in Catfish Production Ponds.

The genus Aeromonas comprises more than 60 recognized species that include many important fish pathogens such as the causative agents of furunculosis and motile Aeromonas septicemia (MAS). Although MAS is typically considered a secondary infection, a new virulent A. hydrophila (vAh) strain has been causing devastating losses to the catfish industry in Alabama since 2009. The objective of this study was to characterize the spatiotemporal distribution of Aeromonas sp. and, specifically, vAh in a commercial catfish farm in western Alabama. We sampled biofilm, sediment, and water from three ponds during four consecutive months during the growing season. Total aerobic counts were between 8.8 × 105 and 1.5 × 106  CFU/mL but were significantly higher in biofilm and sediment than in water throughout the sampling period. Total Aeromonas counts in water samples significantly increased in all three ponds after the month of August and ranged from 7.8 × 103 to 4.9 × 104  CFU/mL. A similar trend was observed in biofilm and sediment samples for which total Aeromonas counts increased in samples taken in late summer to early fall. Over time, the concentration of Aeromonas in water samples decreased by one order of magnitude, while there was a significant increase in sediments as temperature dropped. The virulent vAh was detected in 35.4% of biofilm samples and 22.9% of sediment samples, suggesting that both environments serve as the major reservoir for this pathogen. Future monitoring efforts should focus on targeting sediment and biofilms since samples of these appear to naturally enrich for the presence of vAh and other Aeromonas species.

Bacterial diversity and community structure of the intestinal microbiome of Channel Catfish (Ictalurus punctatus) during ontogenesis.

The acquisition of gut microbes does not occur randomly and is highly dependent on host factors, environmental cues, and self-assembly rules exerted by the microbes themselves. The main objective of this project was to characterize how the gut microbiome develops during the early life stages of Channel Catfish and to identify i) which bacteria are the main constituents of the gut microbiome at different ontogenesis stages, and ii) at which time point(s) the gut microbiome stabilizes. High-throughput Illumina Miseq DNA sequencing of the V4 domain of the 16S rRNA gene was used to assess the microbial community composition during the life stages of Channel Catfish along with water and feed samples. Microbiomes from fertilized eggs, sac fry, swim up fry, pre-fingerlings, and fingerlings were all significantly distinct. OTUs analyses showed that the phylum Proteobacteria, Firmicutes, Fusobacteria and Cyanobacteria dominated the Channel Catfish gut microbiome. During the early stages of ontogenesis, the fish microbiome was dynamic and highly diverse, with significant shifts occurring between fertilized eggs to sac fry (6dph), and from sac fry to swim up fry (15dph). The gut microbiome stabilized between the pre-fingerlings and fingerlings stage (≤90dph) with an observed reduction in species richness. Feed had a more significantly contribution to the microbial colonization of the gut than water. We have identified the period in which the gut microbiome changes rapidly from 15dph until 21dph before stabilizing after 90dph.

Morphological and molecular confirmation of Myxobolus cerebralis myxospores infecting wild­caught and cultured trout in North Carolina (SE USA).

We used microscopy and molecular biology to provide the first documentation of infections of Myxobolus cerebralis (Myxozoa: Myxobolidae), the etiological agent of whirling disease, in trout (Salmonidae) from North Carolina (USA) river basins. A total of 1085 rainbow trout Oncorhynchus mykiss, 696 brown trout Salmo trutta, and 319 brook trout Salvelinus fontinalis from 43 localities across 9 river basins were screened. Myxospores were observed microscopically in pepsin-trypsin digested heads of rainbow and brown trout from the Watauga River Basin. Those infections were confirmed using the prescribed nested polymerase chain reaction (PCR; 18S rDNA), which also detected infections in rainbow, brown, and brook trout from the French Broad River Basin and the Yadkin Pee-Dee River Basin. Myxospores were 9.0-10.0 µm (mean ± SD = 9.6 ± 0.4; N = 119) long, 8.0-10.0 µm (8.8 ± 0.6; 104) wide, and 6.0-7.5 µm (6.9 ± 0.5; 15) thick and had polar capsules 4.0-6.0 µm (5.0 ± 0.5; 104) long, 2.5-3.5 µm (3.1 ± 0.3; 104) wide, and with 5 or 6 polar filament coils. Myxospores from these hosts and rivers were morphologically indistinguishable and molecularly identical, indicating conspecificity, and the resulting 18S rDNA and ITS-1 sequences derived from these myxospores were 99.5-100% and 99.3-99.8% similar, respectively, to published GenBank sequences ascribed to M. cerebralis. This report comprises the first taxonomic circumscription and molecular confirmation of M. cerebralis in the southeastern USA south of Virginia.

Molecular characterization of two opecoelid trematodes from fishes in the Gulf of Mexico, with a description of a new species of Helicometra.

The plagioporine opecoelids Helicometra fasciata (Rudolphi, 1819) Odhner, 1902, and Macvicaria crassigula (Linton, 1910) Bartoli, Bray, and Gibson, 1989 have been reported from fishes in expansive geographic regions, disjointed from their type localities. New material of M. crassigula was collected from near its type locality as well as specimens resembling Helicometra fasciata sensu lato from three triglids in the Gulf of Mexico. Comparisons of the ribosomal DNA (rDNA) sequences, comprising the partial 18S rDNA, internal transcribed spacer region (= ITS1, 5.8S, and ITS2), and partial 28S rDNA gene, from M. crassigula and Helicometra fasciata sensu lato in the Gulf of Mexico were made with sequences deposited in GenBank from those species from the Mediterranean Sea. Results reveal that M. crassigula sensu stricto from the Gulf of Mexico is distinct from the two cryptic species of M. crassigula sensu lato from the Mediterranean Sea and Helicometra fasciata sensu lato in this study differs from H. fasciata sequences from the Mediterranean Sea, thus Helicometra manteri sp. nov. is described.

A new species of Trichosomoididae (Nematoda) from skin of red snapper, Lutjanus campechanus (Perciformes: Lutjanidae), on the Texas-Louisiana shelf, northern Gulf of Mexico.

Eggs and larvae of Huffmanela oleumimica n. sp. infect red snapper, Lutjanus campechanus (Poey, 1860), were collected from the Texas-Louisiana Shelf (28°16'36.58″N, 93°03'51.08″W) and are herein described using light and scanning electron microscopy. Eggs in skin comprised fields (1-5 × 1-12 mm; 250 eggs/mm(2)) of variously oriented eggs deposited in dense patches or in scribble-like tracks. Eggs had clear (larvae indistinct, principally vitelline material), amber (developing larvae present) or brown (fully developed larvae present; little, or no, vitelline material) shells and measured 46-54 µm (x = 50; SD ± 1.6; n = 213) long, 23-33 (27 ± 1.4; 213) wide, 2-3 (3 ± 0.5; 213) in eggshell thickness, 18-25 (21 ± 1.1; 213) in vitelline mass width, and 36-42 (39 ± 1.1; 213) in vitelline mass length with protruding polar plugs 5-9 (7 ± 0.6; 213) long and 5-8 (6 ± 0.5; 213) wide. Fully developed larvae were 160-201 (176 ± 7.9) long and 7-8 (7 ± 0.5) wide, had transverse cuticular ridges, and were emerging from some eggs within and beneath epidermis. The new species differs from its congeners by having eggs <65 µm in total length and that have a brown eggshell when fully developed, an envelope throughout development, and irregularly-dispersed eggshell spines plus a larva >110 µm long with transverse cuticular ridges. The eggs lack a spindle-shaped envelope, polar filaments, and eggshell ridges. This is the first report of a species of Huffmanela from a snapper (Lutjanidae) or from the Gulf of Mexico. A table of egg and larval characteristics, hosts, and localities for Huffmanela spp. is provided.