Huffmanela cf. huffmani (Nematoda: Trichosomoididae) infecting swim bladder, peritoneum, and gonad of variable platyfish, Xiphophorus variatus (Cyprinodontiformes: Poeciliidae) and eastern mosquitofish, Gambusia holbrooki (Poeciliidae) in Florida; taxonomy, phylogenetic analysis, and pathological changes.

Variable platyfish, Xiphophorus variatus (Meek, 1904) (Cyprinodontiformes: Poeciliidae) and eastern mosquitofish, Gambusia holbrooki Girard, 1859 (Poeciliidae) from earthen ponds in west central Florida were examined for parasitic infections. At necropsy, we observed myriad nematodes (adults and eggs), which we identified as Huffmanela cf. huffmani, infecting the swim bladder, gonad, and visceral peritoneum. Nucleotide sequences (small subunit ribosomal DNA, 18S) of H. cf. huffmani from variable platyfish and eastern mosquitofish were identical; likewise for newly obtained 18S sequences of Huffmanela huffmani Moravec, 1987 from the swim bladder of red breast sunfish, Lepomis auritus (Linnaeus, 1758) (Centrarchiformes: Centrarchidae) and warmouth, Lepomis gulosus (Cuvier, 1829) from the San Marcos River (type locality for Huffmanela huffmani Moravec, 1987), Texas. The sequences of H. huffmani and H. cf. huffmani differed by 7 (1%) nucleotides. Pathological changes comprised proliferation of the tunica externa of the swim bladder in low-intensity infections in addition to inflammation, proliferation, and tissue necrosis of swim bladder, peritoneum, and gonad in high-intensity infections. The lesion was severe, affecting the cellular constituents of the swim bladder wall and reducing the size of the swim bladder lumen; potentially reducing swim bladder physiological efficiency. The present study is the first record of a freshwater species of Huffmanela Moravec, 1987 from beyond the San Marcos River, first record of a species of Huffmanela from a livebearer, first nucleotide sequences and phylogenetic analysis for Huffmanela, and first evidence that an infection by a species of Huffmanela causes pathological changes that could impact organ function.

The life cycle of Huffmanela huffmani Moravec, 1987(Nematoda: Trichosomoididae), an endemic marine-relict parasite of Centrarchidae from a Central Texas spring.

The life cycle of the swim bladder nematode Huffmanela huffmani Moravec, 1987 (Trichinelloidea: Trichosomoididae), an endemic parasite of centrarchid fishes in the upper spring run of the San Marcos River in Hays County, Texas, USA, was experimentally completed. The amphipods Hyalella cf. azteca (Saussure), Hyalella sp. and Gammarus sp. were successfully infected with larvated eggs of Huffmanela huffmani. After ingestion of eggs of H. huffmani by experimental amphipods, the first-stage larvae hatch from their eggshells and penetrate through the digestive tract to the hemocoel of the amphipod. Within about 5 days in the hemocoel of the experimental amphipods at 22 °C, the larvae presumably attained the second larval stage and were infective for the experimental centrarchid definitive hosts, Lepomis spp. The minimum incubation period before adult nematodes began laying eggs in the swim bladders of the definitive hosts was found to be about 7.5 months at 22 °C. This is the first experimentally completed life cycle within the Huffmanelinae.

Redescriptions of Spinitectus acipenseri and S. micracanthus (Nematoda, Cystidicolidae), with notes on the taxonomy of Spinitectus-like nematodes parasitising North American fishes.

Based on light microscopical and scanning electron microscopical (SEM) examinations, two North American species of Spinitectus Fourment, 1884, S. acipenseri Choudhury & Dick, 1992 and S. micracanthus Christian, 1972 (Nematoda, Cystidicolidae) are redescribed from museum voucher specimens (S. acipenseri) and those newly collected from centrarchid and some other fishes in the Upper San Marcos River in Texas and the Santee River in South Carolina, USA. The first use of SEM to study S. acipenseri, a parasite of lake sturgeon Acipenser fulvescens Rafinesque (Acipenseridae) in Canada, made it possible to describe dorsal and ventral lips, amphids and sublabia, and the presence of a dorsal barb on the right spicule, which was confirmed to be the most characteristic feature of this species. The SEM study of S. micracanthus, a parasite mainly of centrarchids, enabled us to correctly determine the location of the excretory pore in relation to rings of cuticular spines in the male, and to describe the exact structure of the tip of the male tail, sublabia, phasmids and the presence of a median ventral protuberance on the male tail. Some taxonomic problems of North American species of Spinitectus are discussed. Filaria serrata Linton, 1901 is considered a junior synonym of S. oviflagellis Fourment, 1884. To date, there are 13 valid species of Spinitectus parasitising fishes in North America. Keys to species of Spinitectus-like nematodes from fishes in North American waters are provided.

Title: Redescriptions de Spinitectus acipenseri et S. micracanthus (Nematoda, Cystidicolidae), et notes sur la taxonomie des nématodes de type Spinitectus parasitant les poissons nord-américains. Abstract: Deux espèces nord-américaines de Spinitectus Fourment, 1884, S. acipenseri Choudhury & Dick, 1992 et S. micracanthus Christian, 1972 (Nematoda, Cystidicolidae) sont redécrites à partir de spécimens de musée et d'autres nouvellement collectés aux USA dans les rivières Upper San Marcos (Texas) and Santee (Caroline du Sud), avec l'aide de la microscopie optique et électronique à balayage (MEB). L'utilisation pour la première fois du MEB pour étudier S. acipenseri, un parasite de l'esturgeon Acipenser fulvescens Rafinesque (Acipenseridae) au Canada, a permis de décrire les lèvres dorsales and ventrales, les amphides et sublabia, ainsi que la barbe dorsale sur le spicule droit que nous confirmons être le critère le plus caractéristique de cette espèce. L'étude au MEB de S. micracanthus, un parasite principalement de centrarchides, nous a permis de déterminer correctement la place du pore excréteur en relation avec les couronnes d'épines cuticulaires chez le mâle, ainsi que de décrire la structure exacte de l'extrémité caudale du mâle, des sublabia, des phasmides et la présence d'une protubérance médiane et ventrale sur la queue du mâle. Des problèmes taxonomiques sur les espèces nord-américaines de Spinitectus sont discutés. Filaria serrata Linton, 1901 est considéré synonyme plus récent de S. oviflagellis Fourment, 1884. À ce jour, il y a 13 espèces valides de Spinitectus parasites de poissons en Amérique du Nord. Des clés des espèces des nématodes de type Spinitectus provenant des poissons des eaux nord-américaines sont fournies.

Huffmanela selachii n. sp. (Nematoda: Trichosomoididae: Huffmanelinae): A new species infecting the skin of the great hammerhead shark (Sphyrna mokarran) and the blacktip reef shark (Carcharhinus melanopterus) in the Arabian Gulf, off-shore Saudi Arabia.

From January 2017 - December 2019, 75 out of 850 (8.8 %) great hammerhead sharks from the Arabian Gulf had skin lesions of black irregular discolorations on the ventral surface of the head. The lesions consisted of pencil-like lineations often advancing forward by about 2 mm in back-and-forth looped scribbles often forming a relatively linear bands of about 5-7 cm wide. Similar lesions were also found in the blacktip reef shark from the same area within the same period, and consisted of straight to irregular black lines, extended indiscriminately across the skin of the sharks. Microscopic examination of the skin revealed the presence of dark-brown eggs exhibiting the spindle or ellipsoidal eggs characteristic of Huffmanela sp. The morphometrics of eggs from both hosts were similar (62.9-89.9 µm long and 29.3-56.1 µm wide). The eggshells were smooth with polar plugs protruding or not, with an abruptly truncated crown-like or shoulder-like collar surrounding the plug. The eggs were only found in the epidermal layer of the skin. Based on the unique morphometrics of the eggs, we report a new species, named: Huffmanela selachii n. sp.. This appears to be the first report of Huffmanela from either the great hammerhead shark or the blacktip reef shark, and the third reported Huffmanela in sharks from the Arabian Gulf. It is also one of few species reported from connecting waters of the greater Indian Ocean. This new finding contributes to our understanding of the diversity and ubiquity of Huffmanela sp. in marine creatures.

Geographic isolation facilitates the evolution of reproductive isolation and morphological divergence.

Geographic isolation is known to contribute to divergent evolution, resulting in unique phenotypes. Oftentimes morphologically distinct populations are found to be interfertile while reproductive isolation is found to exist within nominal morphological species revealing the existence of cryptic species. These disparities can be difficult to predict or explain especially when they do not reflect an inferred history of common ancestry which suggests that environmental factors affect the nature of ecological divergence. A series of laboratory experiments and observational studies were used to address what role biogeographic factors may play in the ecological divergence of Hyalella amphipods. It was found that geographic isolation plays a key role in the evolution of reproductive isolation and divergent morphology and that divergence cannot be explained by molecular genetic variation.

The aquatic annelid fauna of the San Marcos River headsprings, Hays County, Texas.

The San Marcos River in Central Texas has been well studied and has been demonstrated to be remarkably specious. Prior to the present study, research on free-living invertebrates in the San Marcos River only dealt with hard bodied taxa with the exception of the report of one gastrotrich, and one subterranean platyhelminth that only incidentally occurs in the head spring outflows. The remainder of the soft-bodied metazoan fauna that inhabit the San Marcos River had never been studied. Our study surveyed the annelid fauna and some other soft-bodied invertebrates of the San Marcos River headsprings. At least four species of Hirudinida, two species of Aphanoneura, one species of Branchiobdellida, and 11 (possibly 13) species of oligochaetous clitellates were collected. Other vermiform taxa collected included at least three species of Turbellaria and one species of Nemertea. We provide the results of the first survey of the aquatic annelid fauna of the San Marcos Springs, along with a dichotomous key to these annelids that includes photos of some representative specimens, and line drawings to elucidate potentially confusing diagnostic structures.

Observations on the distribution and biology of Huffmanela huffmani (Nematoda: Trichosomoididae).

The nematode parasite Huffmanela huffmani Moravec, 1987 (Trichosomoididae) infects swimbladders of fishes in the family Centrarchidae. Only fish collected from the upper San Marcos River (Texas) have been found infected with H. huffmani eggs with a prevalence of 90%. Hundreds of thousands of H. huffmani eggs have been observed in these fish but only a few specimens of adult worms have ever been found. The San Marcos River arises from springs along the Balcones Fault Zone in San Marcos, Hays County, Texas. The restriction of the parasite to the upper San Marcos River and the high prevalence of the parasite eggs in centrarchids would seem to enable one to solve the life cycle of H. huffmani but this has proved false. Here, the insights and experiments used to help define some of the aspects concerning the life cycle of this enigmatic parasite are described. This study of H. huffmani includes a description of the habitat, the known limits of geographic distribution of the parasite, possible dispersal processes, egg characteristics, the testing of a possible intermediate host, Palaemonetes antrorum (Benedict) (Decapoda: Palaemonidae), and the effects of the digestion process on H. huffmani eggs.

Metacercarial distribution of Centrocestus formosanus among fish hosts in the Guadalupe River drainage of Texas.

We examined the gills of wild fish collected from central Texas for Centrocestus formosanus metacercariae to determine whether this temperature-restricted parasite had invaded the thermally dynamic Guadalupe River via an introduced population in its thermally stable tributary, the Comal River. We collected fish from three sites in the Guadalupe River near its confluence with the Comal River (upstream, at, and downstream) and one site in the Comal River. Centrocestus formosanus infected 14 of the 25 species examined (56.0%) and 171 of the individual fish (27.1%). Several of the infected fish represent new host records for the parasite, and two are listed as species of special concern by the state of Texas. Mean metacercarial intensities varied from 8 to 616 among species, and the highest recorded intensity was greater than 800 in two Guadalupe roundnose minnow Dionda nigrotaeniata. Among the 24 species examined from the Guadalupe River, 11 (45.8%) were infected with C. formosanus. Thorough surveys at the study sites yielded no living specimens of the first obligate intermediate snail host (red-rim melania Melanoides tuberculatus), which must be present to perpetuate the parasite. Thus, the infections were probably due to drifting cercariae that had been shed into the water column upstream of the study area in the Comal River. We therefore investigated spatial patterns in cercarial acquisition using caged fish to determine whether drifting cercariae were present in the water column at the study sites. Of 57 uninfected blacktail shiners Cyprinella venusta exposed to Guadalupe River water downstream from and at the confluence, 52 (91.2%) became infected with C. Formosanus metacercariae at a mean rate of 4 metacercariae/d. This finding extends the known geographic range of this invasive exotic parasite and is the first report of the life cycle being advanced in the fish assemblage of a thermally variable temperate stream in the USA.

First record of Bothriocephalus acheilognathi in the Rio Grande with comparative analysis of ITS2 and V4-18S rRNA gene sequences.

Bothriocephalus acheilognathi is an introduced tapeworm in North America often reported as a serious ecological threat to native fishes. In this paper, we report the first record of B. acheilognathi in the Big Bend region of the Rio Grande in Texas (known as the Río Bravo del Norte in Mexico). Identification of B. acheilognathi was confirmed by morphologic and genetic techniques (sequences of ITS2 and V4-18S rRNA genes). Its prevalence was 27% and its intensity ranged from 1 to 5 individuals in a January 2006 collection of 115 red shiners Cyprinella lutrensis. In addition, it was found in the Tamaulipas shiner Notropis braytoni, a Rio Grande endemic and a new host record. The occurrence of B. acheilognathi might have negative ecological impacts on endemic fishes in the Rio Grande. Several of the fishes that could serve as definitive hosts are of conservation concern. Its occurrence also might affect the success of reintroducing the Rio Grande silvery minnow Hybognathus amarus, which is federally listed as endangered, in this portion of the Rio Grande.