Communications: Isolation of Several Mycobacterium Species from Fish

Abstract

Several species of marine fish caught in the wild and of freshwater ornamental fish were used in this study. Infected organs (liver, spleen, and kidney) were sampled for mycobacteria. Decontaminated tissue samples were plated onto selective media for mycobacterial recovery. After initial isolation, fluorescent and acid-fast staining techniques identified bacterial colonies to genus. Profiles of biochemical growth characteristics were used to further identify the isolates to species. Five species of Mycobacterium were identified: M. simiae, M. scrofulaceum, M. marinum, M. chelonae, and M. fortuitum. Of these, M. simiae and M. scrofulaceum have not been previously reported from fish. Tissue samples containing focal granulomatous lesions were prepared for electron microscopic examination.     

New Host and Locality Records for Copepod and Isopod Parasites of Colombian Marine Fishes

Abstract

Two copepods and three isopods, representing five families of Crustacea, are noted from ten hosts, eight of these representing new host records. Cardiodectes rubosus (Copepoda: Pennellidae) is reported from the Atlantic Ocean for the first time. Excorallana tricornis (Isopoda: Corallanidae) may be a more important fish parasite than previously thought.     

Phenotypic and Genotypic Heterogeneity among Streptococcus iniae Isolates Recovered from Cultured and Wild Fish in North America,Central America and the Caribbean Islands

Abstract

Streptococcus iniae, the etiological agent of streptococcosis in fish, is an important pathogen of cultured and wild fish worldwide. During the last decade outbreaks of streptococcosis have occurred in a wide range of cultured and wild fish in the Americas and Caribbean islands. To gain a better understanding of the epizootiology of S. iniae in the western hemisphere, over 30 S. iniae isolates recovered from different fish species and geographic locations were characterized phenotypically and genetically. Species identities were determined biochemically and confirmed by amplification and sequencing of the 16S rRNA gene. Repetitive-element palindromic PCR fingerprinting as well as biochemical and antimicrobial susceptibility profiles suggest that a single strain of S. iniae was responsible for two different disease outbreaks among reef fishes in the Caribbean, one in 1999 and another in 2008. Interestingly, a majority of the isolates recovered from cultured fish in the Americas were genetically distinct from the Caribbean isolates and exhibited a trend toward higher minimal inhibitory concentration with respect to several antibiotics as well as greater genetic variability. The biological significance of this genetic variability is unclear, but it could have implications for future vaccine development and treatment.

Received April 20, 2014; accepted July 7, 2014     

First account of the metazoan parasite fauna of oilfish Ruvettus pretiosus Cocco, 1829 (Perciformes: Gempylidae) in South African waters

Parasites are an important but neglected component of ecosystems that can be used as indicators of host biology and ecology. In the present study, the metazoan parasite assemblage of Ruvettus pretiosus, an understudied but widely distributed predatory gempylid, caught off South Africa was surveyed. A total of seven parasite taxa, including four new infection records (Bolbosoma capitatum, Rhadinorhynchus sp., Hepatoxylon trichiuri and Anisakis sp.), two new locality records (B. capitatum and Rhipidocotyle sp.) as well as the ectoparasitic copepod Sagum foliaceus and the cestode Tentacularia coryphaenae were recovered from the eight specimens examined.     

Antibiotic Resistance of Aeromonas spp. Isolated from Tropical Fish Imported from Singapore

Abstract

To determine the scope of antibacterial resistance among Aeromonas spp., bacterial cultures were taken from a variety of tropical fish species imported from Singapore. The samples were plated on Rimler-Shotts medium for bacterial isolation and identified with both the API20E and Nonfermenter Test Strip systems. Aeromonas hydrophila, A. sobria, and A. caviae were identified in monomicrobic and concomitant infections. Following identification of bacterial isolates, 11 antimicrobials routinely used in the tropical fish industry and 1 new fluoroquinolone were tested for their effectiveness against Aeromonas spp. with the Kirby-Bauer disk-diffusion technique. Aeromonas sobria proved to be the most resistant, often showing susceptibility to only 3 of 12 test drugs. Aeromonas hydrophila was consistently the least resistant. Based on these results, antimicrobial resistance appears to be a rapidly emerging problem in the pet fish industry.     

Abundance of the Parasitic Copepod Caligus elongatus on Wild Pollock near Commercial Salmonid Net-Pens

Abstract

Between July and December 1993, wild pollock Pollachius virens collected near salmonid net-pens on the Atlantic coast of Maine in the United States were examined for parasitic copepods of the family Caligidae to determine the role of pollock as host reservoirs. Only Caligus elongatus was detected on 1,456 pollock sampled. Most pollock (97.39%) had two or fewer sea lice per fish. A maximum prevalence (percent of fish infested) of 50%, an abundance (average number per fish) of 0.9, and an intensity (average number per infested fish) of L79 were recorded. Only one larval C. elongatus was found. No seasonal or geographic trends in infestation statistics were observed.     

Metacercarial Distribution of Centrocestus formosanus among Fish Hosts in the Guadalupe River Drainage of Texas

Abstract

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.

Received October 18, 2010; accepted April 7, 2011     

Aphanomyces invadans and Ulcerative Mycosis in Estuarine and Freshwater Fish in Florida

Abstract

In the spring of 1998, the Florida Fish and Wildlife Research Institute received numerous reports of lesioned or ulcerated fish primarily from the St. Lucie Estuary on the southeast coast of Florida, an area known since the late 1970s for lesions of the ulcerative mycosis (UM) type. From these and archived reports, as well as others received from different areas of Florida, we documented that diseased specimens had randomly distributed skin ulcers (usually reddened or hemorrhagic) with raised irregular margins and, in some cases, deeply penetrating hyphae in the surrounding muscle tissue. Since 1998, 256 fish (comprising 18 species) with ulcerative lesions (from 15 different locations) were confirmed with hyphae in fresh squash preparation or by histological evaluation. Squash preparations revealed nonseptate, sparsely branching, thick-walled hyphae; histological sections revealed mycotic granulomas in the dermis that occasionally penetrated into the skeletal muscle. These pathological characteristics were consistent with UM caused by the oomycete Aphanomyces invadans in Southeast Asia, Japan, Australia, and the United States. For specific identification, six isolates from ulcerated fish were cultured and prepared for molecular characterization using established diagnostic methods. Ribosomal RNA gene sequence analysis identified three isolates as Aphanomyces invadans, one as the oomycete Achlya bisexualis, and two as the ascomycete Phialemonium dimorphosporum. A more extensive survey of 67 ulcerated skin samples from fish collected between 1998 and 2003 was performed using a polymerase chain reaction assay specific for Aphanomyces invadans. Of these, 26 (38.8%) samples from seven fish species and nine collection locations were positive. Confirmation of UM associated with Aphanomyces invadans represents new host records in Florida for the sheepshead Archosargus probatocephalus, striped mullet Mugil cephalus, white mullet Mugil curema, silver perch Bairdiella chrysoura, black drum Pogonias cromis, largemouth bass Micropterus salmoides, and American shad Alosa sapidissima.     

Morphologic,morphometric and contour shape variations of sagittal otoliths of Lepidorhombus spp. in the Aegean Sea

Otoliths are calcareous anatomical structures in the inner ear of fishes, and they can be used in the discrimination of fish species and stocks due to their species-specific shape. Sagittal otoliths in the Lepidorhombus whiffiagonis and Lepidorhombus boscii, two flatfish species of the same genus distributed in the Aegean Sea, were compared using morphology, morphometry and contour shape analyses. Blind and eyed side otoliths of Lepidorhombus species were evaluated separately due to their morphological features and statistical differences in morphometric measurements (p < 0.05). Four analysis groups were formed: blind side otolith of L. whiffiagonis, eyed side otolith of L. whiffiagonis, blind side otolith of L. boscii and eyed side otolith of L. boscii. Morphometric differences in otolith shape were performed by canonical discriminant analysis, and the first discriminant axis explained 97.4% (Wilks λ = 0.270) of the variance between the groups, and the second axis explained 2.3% (Wilks λ = 0.620). Classification success between Lepidorhombus species based on CDA is 100%, and the overall CDA classification score between groups is 76.5%. The wavelet functions obtained in the contour analysis showed high variability in the anterior, posterior-dorsal, antero-dorsal and ventral regions of the otoliths among these four analysis groups. Consequently, it is thought that this study will contribute to the taxonomic classification of fish with the morphological and morphometric differences in the blind and eyed side otoliths and the analyses made with the high discrimination success obtained in Lepidorhombus species.     

Behavior of Rocinela angustata (Isopoda,Aegidae), an Ectoparasite of Alaskan Marine Fishes

Abstract

Observations were made on the feeding and host selection behavior of the aegid isopod Rocinela angustata. Host susceptibility was assessed by placing isopods into laboratory tanks containing 14 common species of Alaska marine fishes. Five species of Sebastes and Blepsias bilobus were attacked within a few minutes of the introduction. Five species—Enophrys bison, Eumicrotremus orbis, Hexagrammos lagocephalus, Pleuronectes asper, and Pholis laeta—were attacked 10–24 h after introduction of the isopods. Three species—Podothecus acipenserinus, Liparis gibbus, and Lycodes brevipes—were not successfully attacked, even when held in isolation with an isopod for 14 d. Starved isopods isolated with B. bilobus attached to fish within 1 h and remained attached 4–14 d. The isopods reattached 1–4 d after detachment. Isopod digestive tracts cleared within 24 h after detachment.     

Survey of Salmonid Pathogens in Ocean-Caught Fishes in British Columbia,Canada

Abstract

A survey of wild fishes captured around marine net-pen salmon farms and from open waters for certain salmonid pathogens was conducted in the coastal waters of British Columbia. Viral hemorrhagic septicemia virus was detected in Pacific herring Clupea pallasi, shiner perch Cymatogaster aggregata, and threespine sticklebacks Gasterosteus aculeatus. Infectious hematopoietic necrosis (IHN) virus was detected in one Pacific herring (collected well away from the farms) and in tube-snouts Aulorhynchus flavidus and shiner perch collected from a farm experiencing an IHN outbreak. Renibacterium salmoninarum was observed in moribund Pacific hakes Merluccius productus collected from within a net-pen and was also detected in several ocean-caught salmon. Aeromonas salmonicida subsp. salmonicida (typical strain) was isolated from a juvenile chinook salmon Oncorhynchus tshawytscha, whereas the atypical strain of this organism was isolated from a lingcod Ophiodon elongatus. Loma salmonae (Microsporea) was observed in chinook salmon, chum salmon Oncorhynchus keta, coho salmon O. kisutch, sockeye salmon O. nerka, and pink salmon O. gorbuscha, all of which were captured well away from net-pens. Loma spp. (Microsporea) were observed in the gills of shiner perch, lingcod, Pacific tomcod Microgadus proximus, Pacific cod Gadus macrocephalus, walleye pollock Theragra chalcogramma, and sablefish Anoplopoma fimbria; all but the first species represent new hosts for Loma. Epitheliocystis, caused by a chlamydia-like organism, was detected in the gills of chinook salmon, chum salmon, coho salmon, pink salmon, lingcod, Pacific cod, Pacific hakes, Pacific tomcod, walleye pollock, sablefish, shiner perch, Dover soles Microstomus pacificus, Pacific sanddabs Citharichthys sordidus, and various species of rockfish Sebastes spp., most of which represent new host records for this infection.     

Susceptibility of Selected Inland Salmonids to Experimentally Induced Infections with Myxobolus cerebralis,the Causative Agent of Whirling Disease

Abstract

Laboratory exposures to the infectious stages (triactinomyxons) of Myxobolus cerebralis demonstrated a range of susceptibility to whirling disease among four species of inland salmonids. Replicate groups of each species were exposed to two concentrations of triactinomyxons, a low dose (100–200 per fish) and a high dose (1,000–2,000 per fish). Exposed fish were evaluated for clinical signs, for severity of microscopic lesions at 35 d, 2 and 5 months, and for spore concentrations in the head cartilage at 5 months. A standard strain of rainbow trout Oncorhynchus mykiss matched for age served as a susceptible species control. Rainbow trout, westslope cutthroat trout O. clarki lewisi, Yellowstone cutthroat trout O. clarki bouvieri, and bull trout Salvelinus confluentus were susceptible to M. cerebralis infections. Clinical signs, including radical swimming (“whirling”) and black tails, were observed at 7 weeks postexposure among rainbow and cutthroat trout challenged at 3 weeks of age. Clinical signs were rare among bull trout exposed at an age of 4 weeks and absent among rainbow and cutthroat trout exposed at 3 months posthatch. Most rainbow, cutthroat, and bull trout were found to be infected when examined at 5 months postexposure. The most severe microscopic lesions among infected fish at 5 months postexposure were found among rainbow trout. Cutthroat trout had less severe lesions, bull trout had mild infections, and no evidence of infection was found among Arctic grayling Thymallus arcticus. Mean spore concentrations among infected fish correlated with the severity of microscopic lesion scores. Rainbow trout had mean concentrations of spores in head cartilage reaching 10⁶, whereas more resistant species such as bull trout had 10⁴ spores; no spores were found among Arctic grayling at 5 months postexposure.     

Mycobacterial Infection in Laboratory-Maintained Atlantic Menhaden

Abstract

Mycobacteriosis can be a chronic wasting disease found in many species of fish examined from wild-caught, cultured, and aquarium-reared populations. An Atlantic menhaden Brevoortia tyrannus with an open ulcer from a wild-caught, laboratory-maintained population was sampled for microbiology. Mycobacterium spp. was recovered from the ulcer and M. fortuitum complex was recovered from the spleen of this fish. Subsequently, 20 additional fish were subsampled from this population to determine the prevalence of the infection. Bacteriology samples were taken from the brain, liver, and kidney. Spleens were homogenized and plated on media enriched for mycobacterial growth, full necropsies were performed, and samples were taken for histology. Bacteriology results showed 100% of the fish in the subsample were infected with mycobacteria. Three species of mycobacteria were isolated from the spleen tissues: M. marinum, M. fortuitum complex, and M. gordonae. Histology results revealed that granulomas, characteristic of mycobacteriosis, were most prevalent in livers but were also found in spleens, posterior kidneys, and hearts. This report adds Atlantic menhaden to the list of species susceptible to mycobacteria.     

Microbial Pollution in Water and Its Effect on Fish

Abstract

A systematic study of microbial contamination of fish grown in ponds in and around Calcutta was initiated as part of the wetland management project in the city. Ponds were either sewage-fed or conventional (i.e., fed by rain and groundwater). Fish from these ponds are used for human consumption, and therefore a quantitative analysis of their microbial content is necessary. When concentrations of Escherichia coli, Salmonelleae (unidentified species), and Staphylococcus aureus in water and different organs of fish from sewage-fed and conventional ponds were compared, water and fish organs from conventional ponds contained about two orders of magnitude more bacterial cells. We tested for Salmonelleae and S. aureus, in addition to E. coli, because of persistent reports from Calcutta hospitals that most enteric and other infectious diseases in this region are caused by these bacteria. Significant linear correlations were found between concentrations of these bacteria in pond water and their recovery from several tissues of the fish.     

Ixodid ticks on domestic dogs in the Northern Cape Province of South Africa and in Namibia

The objective of this study was to determine the species composition of ixodid ticks infesting domestic dogs in the northwestern region of the Northern Cape Province of South Africa and in Namibia. Ticks were collected from February 2008 to January 2009 from dogs presented for a variety of reasons at a veterinary clinic in the Northern Cape Province and at 3 clinics in Namibia. The ticks collected at each place were pooled separately for each month at each locality. Eleven ixodid tick species were collected from dogs in the Northern Cape Province and new locality records for Haemaphysalis colesbergensis and Ixodes rubicundus, new locality and host records for Hyalomma glabrum, and a new host record for Rhipicephalus neumanni are reported. Six tick species were collected from dogs at the 3 clinics in Namibia. The most numerous species on dogs in both countries was R. sanguineus. The present results increase the total number of ixodid tick species collected from dogs in South Africa from 25 to 28.     

Isolation of Acholeplasma laidlawii from Centrarchids in a Central Florida Lake

Abstract

In 1991, the poor physical condition of largemouth bass Micropterus salmoides from Lake Harris, Florida, was associated with the decline of the lake's fishery. The swim bladders of emaciated bass had mild inflammation and ecchymotic hemorrhages. A mycoplasma-like organism isolated from swim bladders was initially believed to be the causative agent. The organism was later identified as Acholeplasma laidlawii by using a fluorescent antibody procedure and was demonstrated to be nonpathogenic. Parenteral injection of the organism into healthy largemouth bass fingerlings produced no signs of disease or difference in growth rate compared with control fish during a 16-month period. Field studies resulted in isolation of A. laidlawii from black crappies Pomoxis nigromaculatus, bluegills Lepomis macrochirus, and redear sunfish L. microlophus, but not from noncentrarchids in Lake Harris or from any fish species in a control fishery (Lake Holly, Florida). The absence of organisms in all emaciated bass, our inability to reproduce the disease, and isolation of the organism from seemingly healthy fish suggest this organism was not pathogenic.     

Introduction,establishment and spread of the Southern mouthbrooder Pseudocrenilabrus philander in the Baakens River,Eastern Cape,South Africa

This paper provides evidence for the extralimital establishment and spread of Pseudocrenilabrus philander in the Baakens River, Eastern Cape, South Africa. Since the first occurrence record at a single locality in 1994 this species has now spread throughout the river and the population contains both reproductive adults and juveniles. Gut content analysis demonstrated potential dietary overlap with native fishes and predation on fish larvae. Pseudocrenilabrus philander in the Baakens River therefore satisfies all criteria for an invasive species in this river and was categorised as invasive using a unified framework for biological invasions.     

Detection of Infectious Pancreatic Necrosis Virus from the Leeches Hemiclepsis marginata and Hirudo medicinalis

Leeches have been reported to harbor several important fish pathogens, including spring viremia of carp virus, infectious hematopoietic necrosis virus (IHNV), and viral hemorrhagic septicemia virus (VHSV), and also may contain blood protozoa. In the present study, leeches were collected from water bodies located in Kurdistan province, Iran. The specimens were tested for IHNV, VHSV, and infectious pancreatic necrosis virus (IPNV) using the PCR method. The results showed that two different species of leeches, Hemiclepsis marginata and Hirudo medicinalis, were infected by IPNV among the seven species studied. The infected leeches were found in areas that were polluted with untreated sewage coming from upstream fish farms culturing Rainbow Trout Oncorhynchus mykiss. In addition, the fish at fish farms in the vicinity had been infected with IPNV 9 months previously. Our results showed that the virus causing infectious pancreatic necrosis is present in the leeches H. marginata and H. medicinalis, suggesting that leeches are a potential source of IPNV in fish farms.

Received October 14, 2015; accepted June 1, 2016 Published online September 29, 2016     

Relationship between Length of Roundtail Chub and Infection Intensity of Asian Fish Tapeworm Bothriocephalus acheilognathi

Abstract

Nonnative parasites have been blamed for the decline of native fish species in the American Southwest. The Asian fish tapeworm Bothriocephalus acheilognathi has parasitized many native fish species, but little is known of its effects on native cyprinid fish hosts. I found a strong negative correlation (Spearman's rank correlation, r_s = ?0.846; P < 0.001) between total length of roundtail chub Gila robusta and number of Asian fish tapeworms. A significant, but weaker negative correlation existed between weight of fish and number of tapeworms (r_s = ?0.687; P < 0.001). In addition, infected fish were significantly shorter (P = 0.0241) than uninfected fish. Infection by Asian fish tapeworm may slow growth, increase susceptibility to infestation, and increase predation on roundtail chub and possibly other native cyprinids.