Vector specificity of Trypanosoma catostomi and its infectivity to freshwater fishes.

Trypanosoma catostomi was found in 36.2% of 558 white suckers (Catostomus commersoni) from Ontario, Canada. The abundance of Actinobdella inequiannulata was 35% (68 leeches/197 suckers examined for leeches). The susceptibility of 3 species of leeches (Hemiclepsis marginata, Desserobdella phalera, and A. inequiannulata) and 7 species of fishes (C. commersoni, Amia calva, Anguilla rostrata, Ictalurus nebulosus, Oncorhynchus mykiss, Perca flavescens, and Esox lucius) to infection with T. catostomi was examined. Metatrypanosomes were found in the crop and proboscis sheath of 13 of 21 A. inequiannulata and in the crop of 10 of 12 H. marginata and 1 of 21 D. phalera. Only flagellates from A. inequiannulata were infective to C. commersoni. Cultured T. catostomi infected C. commersoni and A. calva but not any other fish species. Laboratory-reared C. commersoni were more susceptible than wild-caught specimens. Cultured Trypanosoma phaleri did not infect its natural host, A. calva. Host specificity should be established experimentally before a specific diagnosis is made. Cultures may be useful in simulating factors that affect development in the vector.     

Molecular cloning of cDNA encoding APC11, a catalytic component of anaphase-promoting-complex (APC/C), from goldfish (Carassius auratus), and establishment of in vitro ubiquitinating system

Destruction of cyclin B is required for exit from mitosis and meiosis. A cyclin-degrading system, including anaphase-promoting-complex/cyclosome (APC/C), has been shown to be responsible for cyclin B destruction. Here we present the cloning, sequencing, and expression analysis of goldfish (Carassius auratus) APC11, which encodes the catalytic component of APC/C from goldfish ovary. The cloned cDNA is 348 bp long and encodes 88 amino acids. The deduced amino acid sequence is highly homologous to APC11 from other species. The expression of mRNA for APC11 was ubiquitous among tissues, as opposed to that of mRNA for E2-C, which occurred at a very high level in the ovary. Recombinant goldfish APC11 possesses ubiquitinating activity against cyclin B. We established an in vitro ubiquitinating system of proteins composed of purified recombinant E1, E2-C, and APC11 from goldfish. The reconstructed system for these ubiquitinating enzymes makes it feasible to elucidate the molecular mechanism of cyclin B degradation.     

Temperature tolerance of some estuarine fishes

The temperature tolerance of estuarine fishes Etroplus suratensis (Bloch), Therapon jarbua (Forsskal), Ambassis commersoni (Cuvier) collected from Vellar estuary, South India in 1986-87 were determined in the laboratory. E. suratensis and T. jarbua were found to be more tolerant to a wide range of temperature (16.5-41.5 degrees C for E. suratensis and 13.5-40.6 degrees C for T. jarbua), A. commersoni could tolerate from 15.5 to 38.5 degrees C. The tolerance area was found to be 512 units in E. suratensis, 629 units in T. jarbua and 442 units in A. commersoni. Among the fishes tested T. jarbua had a high tolerance area (629 units) than other fishes tested. It is evident from the results that 15 degrees C increase in acclimation temperature cause a shift of 4.02 degrees C (E. suratensis) and 3.05 degrees C (T. jarbua and A. commersoni) in upper incipient lethal temperature.     

Molecular cloning of cDNA encoding a cyclin-selective ubiquitin carrier protein (E2-C) from Carassius auratus (goldfish) and expression analysis of the cloned gene

Destruction of cyclin B is required for exit from mitosis and meiosis. A cyclin-specific ubiquitinating system, including cyclin-selective ubiquitin carrier protein (E2-C), is thought to be responsible for cyclin B destruction. Here we present the cloning, sequencing and expression analysis of goldfish, Carassius auratus, E2-C which encodes the cyclin-selective ubiquitin carrier protein from goldfish ovary. The cloned cDNA is 677 bp long and encodes 172 amino acids. The deduced amino acid sequence is highly homologous to E2-C from other species. Recombinant goldfish E2-C possesses ubiquitinating activity against cyclin B. The expression of mRNA for E2-C was similar to that of mRNA for cyclin B, occurring at very high level in the ovary. The similarity of the expression pattern of E2-C and cyclin B suggests that E2-C mediates a cyclin-specific ubiquitination.     

History repeated: recent and historical mitochondrial introgression between the current darter Etheostoma uniporum and rainbow darter Etheostoma caeruleum (Teleostei: Percidae)

Incongruence between recognized taxonomy and phylogenetic relationships between two species from a diverse clade (Percidae: Etheostomatinae) of stream fishes was found in a mitochondrial (mt) DNA gene tree. Two darters in subgenus Oligocephalus , Etheostoma uniporum current darter and Etheostoma caeruleum rainbow darter were sampled throughout their sympatric distribution in the Ozark Highlands of the central United States. Sequences from cytochrome (cyt) b and the first intron of the nuclear marker S7 were analysed separately using maximum parsimony and Bayesian methods. Cyt b recovered both species as polyphyletic; E. uniporum haplotypes were interspersed within E. caeruleum . However, both species were monophyletic and non-sister taxa based on S7. The cyt b gene tree pattern is caused by introgressive hybridization resulting in the mtDNA replacement of E. uniporum haplotypes by those of E. caeruleum . Some E. uniporum haplotypes are shared with geographically proximate E. caeruleum , and this is consistent with recent hybridization, while other E. uniporum haplotypes indicate historical sorting of introgressed lineages. The mechanism of introgression is likely asymmetric sneaking behaviour by male E. uniporum , a mating tactic observed in related species. MtDNA replacement may have occurred in E. uniporum due to drift fixation in a historically small female effective population. Additional evidence for darter hybridization will likely be discovered in future molecular genetic surveys of the nearly 200 species in eastern North America.     

Two genotypic groups of morphologically similar fish trypanosomes from the Okavango Delta, Botswana

Blood smears and blood lysate samples from freshwater fishes captured in the Okavango Delta, Botswana, were examined to determine whether their trypanosomes were all Trypanosoma mukasai, a species of supposed broad host specificity and widespread existence across Africa. Trypanosomes and/or babesiosomes occurred in 20/32 blood smears, and morphometric analysis of trypanosomes from 13/32 smears showed features suggestive of T. mukasai, including nuclear indices consistently >1. In 16/32 blood lysate samples from which DNA was extracted, trypanosome DNA was detected in 12/16 by PCR (polymerase chain reaction), using trypanosome-specific ssu rRNA gene primers. Two samples positive for trypanosomes in blood smears yielded no amplifiable trypanosome DNA, but 4 samples with no detectable infection in blood smears were positive for trypanosome DNA, suggesting an overall trypanosome prevalence rate of 17/32 (53%) among fishes and demonstrating the value of PCR in trypanosome recognition. Cloning and sequencing of the 12 amplified fragments revealed 2 genotypic groups among these fish trypanosomes. Group 1 trypanosomes were from cichlids and 3 families of catfishes, Group 2 from 2 types of catfishes. Sequence comparison showed that the consensus Group 1 sequence was most similar to that of Trypanosoma cobitis, representing European fish trypanosomes of the carassii type, while the consensus Group 2 sequence showed similarity with a trypanosome sequence from another African catfish, Clarias angolensis. It was concluded that the identification of T. mukasai remains a problem, but at least 2 genotypic groups of trypanosomes occur in Okavango Delta fishes, and catfishes in this region appear to contain both types.     

Molecular cloning of cDNA encoding a ubiquitin-activating enzyme (E1) from goldfish (Carassius auratus) and expression analysis of the cloned gene

Destruction of cyclin B is required to the mitotic and meiotic cycles. A cyclin-specific ubiquitinating system, including ubiquitin-activating enzyme (E1), is thought to be responsible for cyclin B destruction. Here we present the cloning, sequencing and expression analysis of goldfish, Carassius auratus, E1 from goldfish ovary. The cloned cDNA is 4069 bp long and encodes 1059 amino acids. The deduced amino acid sequence is highly homologous to E1 from other species. Recombinant goldfish E1 could transfer ubiquitin to cyclin-selective ubiquitin-conjugating enzyme. Tissue distribution revealed a single 4.0-kb message ubiquitous among tissues.     

Toxoplasma gondii does not persist in goldfish (Carassius auratus)

Recent reports of toxoplasmosis in marine mammals raise concern that cold-blooded marine animals are a potential source of Toxoplasma gondii infection. To examine the transmissibility of T. gondii to fish, we observed the development of T. gondii tachyzoites inoculated into oviduct epithelial cells of goldfish (Carassius auratus) microscopically in vitro. Further, the survival period of tachyzoites inoculated into goldfish muscle was bioassayed in mice and through PCR analysis. In cell cultures at 37 C, both RH and Beverley strains of T. gondii tachyzoites had penetrated into cells at 6 hr post inoculation, and were multiplying. In cell cultures at 33 C, many tachyzoites of both strains attached to the host cells, but no intracellular tachyzoites were observed at 24 hr post inoculation. In the T. gondii inoculated goldfish kept at 33 C, tachyzoite DNA was detected in the inoculated region on day 3, but not on day 7. When inoculated goldfish were kept at 37 C, live tachyzoites were seen at the inoculation site on day 3, but not on day 7. These results suggest that T. gondii does not persist in fish.     

A BAC library for the goldfish Carassius auratus auratus (Cyprinidae, Cypriniformes)

A goldfish (Carassius auratus auratus) bacterial artificial chromosome genomic library (BAC library) was constructed from one aquarium-bred male specimen (tetraploid, 4n=100, genome size=3.52 pg/cell). The library consists of 128,352 positive clones with an average insert size of 150.4 kb, covering the genome 11-fold. All clones were spotted onto nylon filters and thus are available for screening of genomic regions of interest, such as candidate genes, gene families, or large-sized syntenic DNA regions of cyprinid species. Preliminary screens with two genes were conducted with hybridizing probes to the genes RAG1 and lgi1. RAG1 is a single-copy gene in zebrafish and is duplicated in C. a. auratus. We found a very close correlation between the number of positive BAC clones and the expected library coverage. Two copies of lgi1 were found in zebrafish. We have detected four different copies in C. a. auratus, not in the expected abundance, which indicates some variation in the coverage of the BAC library. The preliminary screens indicate that many duplicated genes that resulted from the ancient fish-specific genome duplication persist in the tetraploid goldfish genome. Hence, the BAC library will provide a useful resource for the future work on comparative genomics, polyploidy, diploidization, and evolutionary genomics in fishes.     

The distribution of the introduced tapeworm Bothriocephalus acheilognathi in Australian freshwater fishes

Native and exotic fishes were collected from 29 sites across coastal and inland New South Wales, Queensland and Victoria, using a range of techniques, to infer the distribution of Bothriocephalus acheilognathi (Cestoda: Pseudophyllidea) and the host species in which it occurs. The distribution of B. acheilognathi was determined by that of its principal host, carp, Cyprinus carpio; it did not occur at sites where carp were not present. The parasite was recorded from all native fish species where the sample size exceeded 30 and which were collected sympatrically with carp: Hypseleotris klunzingeri, Hypseleotris sp. 4, Hypseleotris sp. 5, Phylipnodon grandiceps and Retropinna semoni. Bothriocephalus acheilognathi was also recorded from the exotic fishes Gambusia holbrooki and Carassius auratus. Hypseleotris sp. 4, Hypseleotris sp. 5, P. grandiceps, R. semoni and C. auratus are new host records. The parasite was not recorded from any sites in coastal drainages. The only carp population examined from a coastal drainage (Albert River, south-east Queensland) was also free of infection; those fish had a parasite fauna distinct from that of carp in inland drainages and may represent a separate introduction event. Bothriocephalus acheilognathi has apparently spread along with its carp hosts and is so far restricted to the Murray-Darling Basin. The low host specificity of this parasite is cause for concern given the threatened or endangered nature of some Australian native freshwater fish species. A revised list of definitive hosts of B. acheilognathiis presented.     

The effects of pulp mill effluent on the sex steroid binding protein in white sucker (Catostomus commersoni) and longnose sucker (C catostomus)

The objective of this study was to characterize the effects of pulp mill effluent on the properties of the sex steroid binding protein (SBP) in the plasma of white sucker (Catostomus commersoni) and longnose sucker (C. catostomus). SBPs which specifically bind estradiol and testosterone with high affinity (k(D) approximately 3 nM) and low capacity (B(max) approximately 73-81 nM) were identified in both species. Subsequent studies determined if the properties of the SBP in white sucker exposed to bleached kraft mill effluent (BKME) at Terrace Bay, ON, and in longnose sucker exposed to BKME at Grande Prairie, AB. differed from appropriate reference fish. There were no effects of BKME exposure on the binding affinity (k(D)) of the SBP in either species, but there was a significant increase in the binding capacity (B(max)) of longnose sucker SBP exposed to BKME. The livers of nai;ve white sucker exposed to effluent at Terrace Bay or a bleached sulfite/groundwood mill in Edmundston, NB, rapidly accumulated compounds of differing hydrophobicity that bound to both the white sucker and goldfish (Carassius auratus) SBP. Conversely, there was reduced accumulation of SBP ligands in the bile of effluent-exposed fish. We have demonstrated that constituents present within pulp mill effluent bind to both the white sucker and goldfish SBP, and that native species residing downstream of pulp mill effluents may experience modifications in the properties of their SBP.     

The morphology and systematic status of Khawia rossittensis (Szidat, 1937) and K. parva (Zmeev, 1936) (Cestoda: Caryophyllidea), parasites of cyprinid fishes

The morphology of the two little-known fish cestodes of the genus Khawia Hsü, 1935, K. rossittensis (Szidat, 1937) and K. parva (Zmeev, 1936) from cyprinid fishes, were studied on the basis of newly collected specimens from goldfish Carassius auratus auratus (L.) and museum specimens, respectively. This paper provides the first detailed species diagnosis for K. rossittensis from Slovakia, which is compared with specimens from different geographical regions and K. parva, a somewhat similar Far Eastern species from the same host. The two species differ in scolex morphology, anterior extent of the vitelline follicles, shape of the ovary and size of the eggs. Based on these differences, K. rossittensis and K. parva are considered to be separate taxa. K. parva, listed among the "species incertae sedis" by Mackiewicz (1972) and even within Caryophyllaeus Gmelin, 1790 by Schmidt (1986), should be considered a valid species of Khawia. The results support the previous conclusions of Kulakovskaya (1961), Dubinina (1971) and Protasova et al. (1990).     

Cryptobia cataractae from the blood of Semotilus atromaculatus: structure and division in the fish

A cryptobiid was found in the blood of 2 of 9 Semotilus atromaculatus from a tributary of the Saugeen River in Ontario, Canada. Blood inoculation produced an infection in 2 uninfected S. atromaculatus but not in any Oncorhynchus mykiss, Catostomus commersoni, or Carassius auratus. The flagellate was identified as Cryptobia cataractae, based on host restriction. Cryptobia cataractae occurred as slender and broad forms (body width 3.0-8.7 microns). The length of the anterior flagellum was equal to body length, whereas that of the free recurrent flagellum was half body length. Cryptobia cataractae divided by equal binary fission that produced elongate, slender daughter cells.     

西藏拉鲁湿地和茶巴朗湿地外来鱼类群落结构及变动趋势

研究于2019—2021年在西藏拉鲁湿地和茶巴朗湿地使用刺网和地笼等组合渔具开展了鱼类实地调查, 分析了外来鱼类物种组成、优势种和群落功能多样性, 并结合已发表的文献资料, 探讨了西藏湿地外来鱼类群落结构的变动趋势及影响。结果表明, 共采集到外来鱼类10种, 隶属于4目5科, 包括鲫(Carassius auratus)、麦穗鱼(Pseudorasbora parva)、鲤(Cyprinus carpio)、丁鱥(Tinca tinca)、棒花鱼(Abbottina rivularis)、泥鳅(Misgurnus anguillicaudatus)、大鳞副泥鳅(Paramisgurnus dabryanus)、鲇(Silurus asotus)、小黄?(Micropercops swinhonis)和中华青鳉(Oryzias sinensis)。外来鱼类的个体数和重量分别占本地种和外来种总渔获物的94.86%和70.71%, 其中主要组成为鲫和麦穗鱼等小型鱼类。相对重要性指数(IRI)表明鲫、麦穗鱼、大鳞副泥鳅和鲤4种为拉鲁湿地外来鱼类优势物种, 鲫、麦穗鱼和小黄?3种为茶巴朗湿地的优势物种。在拉鲁湿地和茶巴朗湿地鱼类种类组成、个体数量和生物量中外来鱼类均已占据优势地位, 且外来鱼类的群落功能多样性(FD)亦高于土著鱼类。结合历史数据, 西藏拉鲁湿地和茶巴朗湿地已记录外来鱼类12种, 其中具高度入侵风险物种达6种, 外来鱼类物种数从2000年左右的2种增加到现在的10种, 且丁鱥和中华青鳉分别在西藏自然水体中和茶巴朗湿地中首次被发现。此外, 两个湿地12种外来鱼类均为拉萨市场中的常见经济鱼类及携带鱼类, 是由于放生而进入湿地自然水体中, 且鲫、大鳞副泥鳅和麦穗鱼等均已建立自然种群。因此, 加强对放生的科学管理及开展外来鱼类的长期监测, 是防范外来鱼类入侵及高原湿地生物多样性保护的迫切任务。     

Evolutionary and physiological variation in cardiac troponin C in relation to thermal strategies of fish

Striated muscle contraction is initiated when troponin C (TnC) binds Ca(2+), which activates actinomyosin ATPase. We investigated (i) the variation between cardiac TnC (cTnC) primary structure within teleost fish and (ii) the pattern of TnC expression in response to temperature acclimation. There were few differences between rainbow trout (Oncorhynchus mykiss), yellowfin tuna (Thunnus albacares), yellow perch (Perca flavescens), goldfish (Carassius auratus), white sucker (Catostomus commersoni), and icefish (Chaenocephalus aceratus) in cTnC amino acid sequence. No variation existed in the regulatory Ca(2+)-binding site (site 2). The site 3 and 4 substitutions were limited to residues not directly involved in Ca(2+) coordination. Fish cTnC primary structure was highly conserved between species (93%-98%) and collectively divergent from the highly conserved sequence seen in birds and mammals. Northern blots and polymerase chain reaction showed that thermal acclimation of trout (3 degrees, 18 degrees C) did not alter the TnC isoform pattern. While cardiac and white muscle had the expected isoforms-cTnC and fast troponin C (fTnC), respectively-red muscle unexpectedly expressed primarily ftnC. Cold acclimation did not alter myofibrillar ATPase Ca(2+) sensitivity, but maximal velocity increased by 60%. We found no evidence that TnC variants, arising between species or in response to thermal acclimation, play a major role in mitigating the effects of temperature on contractility of the adult fish heart.     

A comparison of Fusarium avenaceum and Fusarium caeruleum as causes of wastage in stored potato tubers

Fusarium avenaceum is reported for the first time as a cause of rotting of potato tubers in Britain. The progress of rotting in tubers infected with F. avenaceum has been compared with dry rot due to F. caeruleum in the laboratory, clamp and potato store. Of the four varieties, Majestic, King Edward, Doon Star and Arran Pilot, tested for susceptibility, King Edward was the most susceptible to F. avenaceum and Doon Star to F. caeruleum.
Optimum temperatures for growth on potato-dextrose agar were 20-25 C. for F. avenaceum and 20 C. for F. caeruleum ; maximum temperatures were > 30 and 30 C. respectively. For infection of wounded potato tubers, cardinal temperatures for F. avenaceum were similar to those on agar, but for F. caeruleum the optimum was 15 C. and the maximum 25 C. The optimum temperature for rotting tended, with both species, to be higher in the more susceptible potato varieties. At low temperatures F. caeruleum caused quicker rotting than did F. avenaceum , even though its rate of growth on agar was scarcely more than half that of the latter.
High humidity favoured rotting especially by F. avenaceum; F. caeruleum was more tolerant of relatively low humidity. Both species caused quicker rotting in the clamp than in store, even though there was no appreciable difference in mean temperature between the clamp and the store. This was attributed to the higher atmospheric humidity in the clamp.     

Host use evolution in Chrysochus milkweed beetles: evidence from behaviour, population genetics and phylogeny

In two sister species of leaf beetles with overlapping host associations, Chrysochus auratus and C. cobaltinus, we established diet breadth and food preference of local populations for evaluation together with genetic differentiation between populations. While C. auratus turned out to be monophagous on the same plant wherever we collected the beetles, the studied populations of C. cobaltinus fed on three different plant species in the field. Plant preference and ranking of the potential host plants significantly differed between these populations. The amount of genetic differentiation between populations was measured by a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay of a 1300 bp mitochondrial DNA (mtDNA) sequence. In addition, the dominant genotypes of all populations were sequenced. No genetic differentiation between the populations of C. auratus could be detected in the RFLP assay and sequence divergence was low (= 0.3%). In C. cobaltinus, on the other hand, genetic differentiation between populations was high, revealing a lack of gene flow over a much smaller scale and a maximum of 1.3% sequence divergence. C. cobaltinus thereby has the prerequisites for host race formation on different plants from the original host spectrum. Our sequence-based phylogeny estimate allows us to reconstruct historical diet evolution in Chrysochus. Starting from an original association with Asclepiadaceae, the common ancestor of C. auratus and C. cobaltinus included Apocynaceae in its diet. The strict specialization on Apocynum and the loss of acceptance of Asclepiadaceae observed in C. auratus could have resulted from a process similar to that displayed by C. cobaltinus populations.     

Effects of brackish water on growth, feed conversion and energy absorption efficiency by juvenile euryhaline and freshwater stenohaline fishes

Among six species of juvenile fishes (<6 months old), stenohaline species (channel catfish Ictalurus punctatus and goldfish Carassius auratus ) had their highest specific growth rate ( G ) and most efficient food conversion ratio ( E _C) and energy absorption efficiency ( I _E) in fresh water. Three of the euryhaline species (rainbow trout Oncorhynchus mykiss , striped bass Morone saxatilis and Gulf sturgeon Acipenser oxyrinchus desotoi ) had higher G and had more efficient E _C and I _E in 3 and 9‰ salinities than in lower salinities (fresh water and 1‰). For brown trout Salmo trutta (age 3–4 months), 9‰ was above the optimum level for G and E C. However, I _E for brown trout was not significantly different at 3 and 9‰ salinities. Over the salinity range tested, channel catfish had the largest change in G , E _C and I _E, while changes for euryhaline species were relatively small. Although all species tested survived and grew in all treatments, salinities as low as 1‰ adversely affected the stenohaline species, and 9‰ adversely affected brown trout.     

DRY-ROT DISEASE OF THE POTATO

Inoculation of wounded tubers at intervals throughout the storage season showed that, after harvesting, resistance to infection by F. avenaceum was maintained for a longer period than resistance to F. caeruleum , although eventually the tubers became equally susceptible to both fungi. Tubers were less readily infected through clean-cut than through scarified wounds. Resistance to infection was greater when an interval elapsed between wounding and inoculation than when the wound was inoculated immediately; this was related to an increase in the intrinsic ability of the cells near the wound to resist infection rather than to suberization of the wounded surface or wound periderm formation.
Histological studies showed that F. caeruleum grew through the intercellular spaces and that the adjacent host cells remained alive, often for considerable periods, whereas F. avenaceum killed and penetrated the cells with which it came into contact. Restriction of incipient or established lesions caused by F. caeruleum and of incipent lesions caused by F. avenaceum was associated with suberin deposition on the host cell walls and in the intercellular spaces; in established lesions caused by F. avenaceum , restriction was attributed to an increase in the intrinsic resistance of the adjacent host cells, similar to that found near a wounded surface.