镰刀菌对鱼类致病性和组织病理研究

研究采用4种不同的人工感染方式,对模式鱼种金鱼实施镰刀菌(Fusarium spp.)FF08人工感染,以期探讨镰刀菌在不同感染方式下对鱼体的致病强度,同时通过石蜡切片和HE染色进一步研究了其对鱼体组织病理性损伤机制。结果表明:腹腔、肌肉和创伤感染均对金鱼有很强的致死性,15d内死亡率分别达到100%、93%和100%,鱼体均出现与自然发病相同的发病症状。病理分析表明镰刀菌FF08感染可引起鱼体肌肉组织、肝、肾、肠道均出现较严重的损坏。主要表现为肌纤维细胞变性、坏死、溶解,大量炎细胞浸润,并有菌丝侵入肌肉组织;肝细胞水肿、充血、出血、血管发生血栓,并且镰刀菌分生孢子侵入组织,有强烈的细胞炎性反应;肾脏充血、出血、部分组织坏死崩解;肠道堵塞、肠系膜脱落等病理症状。研究将为鱼类镰刀菌病的侵染机制及防治奠定理论基础。       

Ichthyophonus infections in cultured marine fish from Spain

Ichthyophonus sp. is reported for the first time in Mugil capito (thinlip grey mullet) and Li a saliens (leaping grey mullet). The fungus was also found in L. aurata (golden grey mullet), Dicentrarchus labrax (sea bass), Sparus aurata (gilthead sea bream) and Scophthalmus maximus (turbot), whereas Mugil cephalus (grey mullet) was not parasitized. In fish sampled periodically, the highest prevalences were observed in sea bass and the lowest in turbot. Among the fish sampled occasionally, the fungus was found associated to an epizootic in thinlip grey mullet. Ichthyophonus was never found in fish weighing <0·5 g. An increase in the prevalence of infection with the age of turbot and gilthead sea bream was observed. Gilthead sea bream and sea bass showed higher prevalences in a closed system than in open and semi-intensive systems. Multinucleate spherical spores, hyphae and endospores of Ichthyophonus sp. parasitized different organs of thinlip and leaping grey mullets, though infection intensity was maximal in the spleen. In the remaining fish, the fungus was found mainly in the trunk kidney, where it appeared frequently in a necrotic form. Ichthyophonus sp. can be considered a potential threat for marine fish aquaculture, especially in culture conditions which may favour the introduction and transmission of the fungus.     

Infection Characteristics of Clonorchis sinensis Metacercariae in Fish from Republic of Korea

The infection nature of Clonorchis sinensis metacercariae (CsMc) in freshwater fish hosts is closely related to the transmission of human clonorchiasis. This article reviewed the infection characteristics of CsMc in freshwater fish in the Republic of Korea (Korea). The status of CsMc infection was examined in a total of 17,792 cyprinid fish of 49 species in 9 water systems, which included Hantan-/Imjin-gang, Han-gang, Geum-gang, Mangyeong-gang, Yeongsan-gang, Tamjin-gang, Seomjin-gang, Nakdong-gang, and streams in the east coastal areas from 2010 to 2020. The infection status of CsMc was examined according to cyprinid fish species and water systems, after which analyzed by endemicity and susceptibility index. The high endemicity was shown in the cyprinid fish from 3 regions (6.1%) in the upper reaches of Nakdong-gang, such as Banbyeon-cheon (stream), Yongjeon-cheon, and Wi-cheon. The moderate levels were observed in fishes from 8 regions (16.3%), and low endemicity was shown in fishes from 20 regions (40.8%). No CsMc were detected in fish from 18 regions (36.7%). The susceptibility of CsMc in index fish, Puntungia herzi, was found to be a reliable index without examination of other fish species. CsMc infection rates were closely related to subfamily groups in the cyprinid fish hosts in a highly endemic area. In Korea, a total of 58 fish species in 10 families has been listed as the second intermediate hosts for C. sinensis. This review provides several novel features of CsMc infection and clarifies the species of second intermediate freshwater fish host in Korea.     

长江中游长吻鮠、蛇鮈寄生粘孢子虫感染率的季节动态

1.寄生长吻鮠上的粘孢子虫有3种:巨囊两极虫和四极虫一种,寄生在胆囊中,碘泡虫一种寄生在肾脏中;寄生蛇鮈上的粘孢子虫有2种:湖北碘泡虫,寄生在脑、鳃、肾、体腔、肠,主要寄生在脑,楚克拉虫一种,寄生在胆囊中。2.粘孢子虫对长吻鮠、蛇鮈的感染率存在明显的季节变化;对长吻鮠的感染率最高值出现在6月(95%),最低值出现在2月(43%),年平均值为65%;湖北碘泡虫对蛇鮈的总感染率最高值出现在12—2月(63—67%),其次在8月(58%),春秋两季感染率较低(36—44%),年平均值为51%。3.总感染率(IR)与鱼体全长(TL)的关系:长吻鮠IR=0.956-0.009TL;蛇鮈IR=-2.0678+0.3838(TL)-0.0136(TL)2,8厘米以下的蛇鮈未发现感染。4.粘孢子虫的感染率与长吻鮠、蛇鮈的性别无关。5.感染粘孢子虫的长吻鮠,肥满度下降不大(0.04),感染粘孢子虫的蛇鮈,肥满度下降0.12,体重平均下降12.4%。6.长吻鮠中寄生粘孢子虫的总感染率(IR)与长江中游水温(T℃)呈显著正相关,二者关系可用下式描述:IR=0.3129+0.0192(T℃)。蛇鮈寄生粘孢子虫的总感染率(IR)与水温(T℃)之间的关系呈一反抛物线型、两者关系可用下式描述 IR=1.1228-0.073777T+0.001795T2       

A new species, Aphanomyces salsuginosus sp. nov., isolated from ice fish Salangichthys microdon

A species of Aphanomyces was isolated from the ice fish Salangichithys microdon living in brackish water in Japan. White cotton-like growth was found on the heads and fins of the fish. Hyphae penetrated into the dermal layers, subcutaneous tissues, muscular layers, and cartilaginous tissue of the mandible and maxilla; these hyphae were associated with cellular debris and lesions in host tissue. White fluffy colonies from subcultures of these growths were isolated on glucose–yeast agar plates with 0.5% sodium chloride (NaCl). These isolates consisted of delicate, slightly wavy, and moderately branched hyphae. Zoosporangia were isodiametric with the vegetative hyphae. Oogonia were abundant and approximately 21–33 μm in diameter, with irregular short papillae. Generally they were spherical or subspherical and only rarely pyriform. Individual oogonia usually contained a single oospore, which was spherical and 19–27 μm in diameter, with a large shiny vesicle. Antheridial branches, when present, were usually androgynous; however, they were sometimes monoclinous or diclinous. The optimal growth temperature of the isolates was 20°C, and cultures grew well at low salinity (0–0.5% NaCl). Phylogenic analysis based on the internal transcribed space 1-5.8S-ITS 2 of the ribosomal RNA gene indicates that these isolates will be an as-yet unidentified species of Aphanomyces.     

Girdles as the main infection site for Paradeontacylix kampachi (Sanguinicolidae) in the greater amberjack Seriola dumerili

In this study, we provide new information about the habitats selected by the blood fluke Paradeontacylix kampachi in the greater amberjack Seriola dumerili based on an exhaustive anatomical examination. From May to October 1998, 21 fish of the 0+ age class were collected from tanks of the Spanish Institute of Oceanography in Puerto de Mazarrón, Spain, for parasitological analysis. Individuals of P. kampachi were found in 17 of the 21 fish analysed (mean intensity +/- SD: 13.6 +/- 16.6; median: 6). Worms occurred in the girdles, cephalic kidney, sinus venosus, kidney and branchial arteries. A Friedman test with a post-hoc contrast revealed a significantly higher number of worms in the girdles when compared with the other sites, suggesting this may be the main habitat for P. kampachi. This location had never been reported as a habitat for any species of Paradeontacylix, probably because it had not been examined before. Girdles should be routinely examined to estimate the actual intensity of infection and to maximize the likelihood of finding this species of digenean.     

Infection by Parvicapsula sp. (Myxozoa) is associated with mortality in sea-caged Atlantic salmon Salmo salar in northern Norway

In March 2002, 3 seawater farms in northern Norway experienced high mortality among Atlantic salmon postsmolts. A myxosporean parasite assigned to the genus Parvicapsula was detected in the pseudobranchs of diseased fish, and extensive destruction of this organ was observed. The parasite was also found in the gills, liver and kidney of some fish. Based on host species, spore morphology, and the unusual site preference of the parasite, it is likely that it represents a hitherto undescribed species. The diseased fish had been transferred to seawater in September 2001, and it is believed that the infection took place shortly after exposure to seawater. The source of infection is unknown.     

Histopathology of cultured sea bream Sparus aurata infected with sanguinicolid trematodes

The present study is the first report of a sanguinicolid infection affecting sea bream Sparus aurata cultured in net cages in the NE of Spain. The disease was associated with trickling mortalities during the cold season (1999 and 2000). Examination of gill wet mounts of the affected population revealed that sanguinicolid infection was present in 82.6 and 100% of the fish sampled in 1999 and 2000, respectively. Adult flukes, which were located in the kidney, were tentatively identified as members of the family Sanguinicolidae, subfamily Cardicolinae. Eggs and miracidia were found in the gill vascular structures. The inflammatory response triggered by the parasites was moderate and the lesions caused by either eggs and miracidia in the gills or adult flukes in the kidney were not extremely severe, possibly because of the moderate intensity of the parasitosis. Histological observations of sanguinicolid infected sea bream presented here are compared with those reported in other fish species. The role played on sea bream morbility and mortality by other factors (occurrence of a simultaneous moderate monogenean infection, immunological impairement related to low water temperatures) is discussed.     

An intranuclear microsporidium associated with acute anemia in the chinook salmon, Oncorhynchus tshawytscha

An intranuclear microsporidium is described from hemoblastic cells of the chinook salmon, Oncorhynchus tshawytscha. The infection is associated with an acute anemia in the fish. Up to 47% of the hemoblast nuclei were infected in anemic fish. The organisms, found only in spleen and kidney tissues, were 1-2 microns in diameter and consisted of vegetative and early sporulation forms. This microsporidium differs from known species which parasitize fish in its tissue location; however, the absence of mature spores and other life cycle stages precludes determination of its precise taxonomic identity.     

柿树炭疽菌侵染不同柿树种、品种和部位的细胞学特征

用柿树炭疽病菌Colletotrichumgloeosporioides的分生孢子制备孢子悬浮液,接种无核柿、野柿、冬柿和浙江柿的新梢、叶柄和叶片,并观察致病性、附着胞形成和侵染特性。柿树炭疽菌可以侵染无核柿枝条和叶柄以及野柿枝条,但不侵染无核柿叶片、野柿叶柄和叶片,也不侵染冬柿和浙江柿枝条、叶柄和叶片。室内接种试验与田间自然发病结果一致。柿树炭疽菌在不同柿树表面均能形成附着胞,附着胞产生在寄主表皮背斜细胞壁间结合处(JACWs)或近结合处的百分率达81%~93%。接种12h后,不同柿树表面都有附着胞形成;36h后,无核柿枝条、叶柄中有侵染菌丝存在;48h后,无核柿枝条、叶柄中观察到膨大初生菌丝和较细次生菌丝,初生菌丝可扩展到相邻细胞中,而野柿枝条中仅观察到侵染菌丝;60h后,野柿枝条中也观察到膨大的初生菌丝和较细的次生菌丝,但初生菌丝仅局限在最初侵染的细胞中,无核柿枝条和叶柄以及野柿枝条中都有分枝的次生菌丝在细胞内、细胞间或相邻的细胞中扩展;直到接种90h后,在冬柿和浙江柿上都未观察到侵染菌丝的形成。结果表明,柿树炭疽菌在不同柿树种和品种上侵染菌丝的形成和扩展方式可能是其寄主专化性(或致病性)差异的重要机制之一。     

眼斑拟石首鱼虹彩病毒病的诊断

对广东省珠海市每年6—10月池塘养殖的眼斑拟石首鱼(Sciaenop socellatus)暴发不明原因疾病进行了研究。通过对该病的流行病学调查,结合症状观察、病原分离鉴定、病理学变化,结果显示:该病具有明显的传染性,解剖病鱼可见鳃丝贫血、脾脏和肾脏肿大、出血,通过镜检未检测到大量的寄生虫;从发病鱼的肝、脾、肾脏和肠中未分离到致病菌;在病鱼肝、脾和肾组织病理切片中观察到有大量肿大细胞;通过使用OIE 推荐的虹彩病毒特异性引物进行PCR扩增出570 bp左右的目的片段,MCP基因序列比对显示扩增获得的基因序列与真鲷虹彩病毒(RSIV)的基因序列同源性高达99.2%。综合以上结果表明导致眼斑拟石首鱼发病死亡的原因是感染了细胞肿大属虹彩病毒而引起的虹彩病毒病。     

Experimental infection of several fish species with the causative agent of Kuchijirosho (snout ulcer disease) derived from the tiger puffer Takifugu rubripes.

Kuchijirosho (snout ulcer disease) is a fatal epidemic disease which affects the tiger puffer, Takifugu rubripes, a commercial fish species in Japan and Korea. To assess the possibility that non-tiger puffer fish can serve as reservoirs of infection, 5 fish species were challenged by infection with the extracts of Kuchijirosho-affected brains from cultured tiger puffer: grass puffer T. niphobles, fine-patterned puffer T. poecilonotus, panther puffer T. pardalis, red sea bream Pagrus major, and black rockfish Sebastes schlegeli. When slightly irritated, all these species, especially the puffer fish, exhibited typical signs of Kuchijirosho, i.e., erratic swimming, biting together and bellying out (swelling of belly), as generally observed in tiger puffers affected by Kuchijirosho. Although the mortalities of the 2 non-puffer species were lower, injection of the extracts prepared from the brains of both inoculated fish into tiger puffer resulted in death, indicating that the inoculated fish used in this experiment have the potential to be infected with the Kuchijirosho agent. Condensations of nuclei or chromatin in the large nerve cells, which is a major characteristic of Kuchijirosho, were histopathologically observed to some extent in the brains of all kinds of puffer fish species infected. These findings suggest that the virus can spread horizontally among wild and cultured puffers and even among fishes belonging to different orders.     

An Intranuclear Microsporidium Associated with Acute Anemia in the Chinook Salmon,Oncorhynchus tshawytscha1

An intranuclear microsporidium is described from hemoblastic cells of the chinook salmon, Oncorhynchus tshawytscha. The infection is associated with an acute anemia in the fish. Up to 47% of the hemoblast nuclei were infected in anemic fish. The organisms, found only in spleen and kidney tissues, were 1–2 μm in diameter and consisted of vegetative and early sporulation forms. This microsporidium differs from known species which parasitize fish in its tissue location; however, the absence of mature spores and other life cycle stages precludes determination of its precise taxonomic identity.     

Effect of choline deprivation on the growth, morphology and ultrastructure of a choline-requiring mutant of Aspergillus nidulans

Abstract On minimal medium in the absence of added choline, conidiospores of the cho C3 choline-requiring mutant of Aspergillus nidulans germinated and grew normally for approximately 17 h at 37°C. After this period a characteristic aberant morphology was produced by the swelling of small regions at or close to the majority of hyphal apices, creating spherical structures up to 10 μm in diameter from hyphae which were typically 1–2 μm in diameter. During the swelling of these structures, apical extension showed and stopped. This apparently indicated a switch from polarised to unpolarised growth. Electron microscopic examination of ultrathin sections of the swellings indicated that they had thicker walls (158.2 ± 42.1 nm) than the immediately adjacent hyphae (88.0 ± 17.6 nm) and that wall thickness of the swellings increased with increasing age and size. However, the absence of any accumulation of cytoplasmic vesicles within the swellings indicated that they were not simply produced by diversion that they normal polarised flow of vesicles from the tip to the side walls of hyphae.     

Fish infected with Sphaerospora spp. Thélohan (Myxosporea) from waters enzootic for proliferative kidney disease of salmonids

Sphaerospores were found among three species of fish examined from waters known to be enzootic for proliferative kidney disease (PKD) of salmonids. They were detected in the renal tubules of both hatchery-reared rainbow trout (Salmo gairdneri) exposed to the infectious stage of PKD and in chubs (Gila bicolor) in the headwaters of a hatchery where PKD is enzootic. Sticklebacks (Gasterosteus aculeatus) collected near net pens where Pacific salmon had experienced a PKD epizootic were also found to harbor sphaerospores in the lumen of the kidney tubules. The latter two host species contained developmental stages of a myxosporidan in the blood and in the lumen of the kidney tubules which are similar to those of PKX, the causative agent of PKD in salmonid fish. The sphaerospores observed in the rainbow trout are the first to be observed in this species. The similarity to previously observed developmental stages, rarity, and presence of these sphaerospores in salmonid fish from a hatchery where PKD is enzootic suggest that they are the most mature stage of the PKX myxosporidan yet observed.     

Lower filamentation rates of Candida dubliniensis contribute to its lower virulence in comparison with Candida albicans

Candida albicans and C. dubliniensis are very closely related yeast species. In this study, we have conducted a thorough comparison of the ability of the two species to produce hyphae and their virulence in two infection models. Under all induction conditions tested C. albicans consistently produced hyphae more efficiently than C. dubliniensis. In the oral reconstituted human epithelial model, C. dubliniensis isolates grew exclusively in the yeast form, while the C. albicans strains produced abundant hyphae that invaded and caused significant damage to the epithelial tissue. In the oral-intragastric infant mouse infection model, C. dubliniensis strains were more rapidly cleared from the gastrointestinal tract than C. albicans. Immunosuppression of Candida-infected mice caused dissemination to internal organs by both species, but C. albicans was found to be far more effective at dissemination than C. dubliniensis. These data suggest that a major reason for the comparatively low virulence of C. dubliniensis is its lower capacity to produce hyphae.     

Modulation of granulocyte responses in three-spined sticklebacks Gasterosteus aculeatus infected with the tapeworm Schistocephalus solidus

Leukocytes isolated from the head kidney and peripheral blood of 3-spined sticklebacks Gasterosteus aculeatus L. were analysed by means of flow cytometry during infection with the tapeworm Schistocephalus solidus (Müller, 1776). Although parasites increased their body weight continuously throughout the observation period (98 d), proportions of granulocytes increased in blood and head kidney only up to Day 63 post-infection (p.i.). Thereafter, declining proportions of granulocytes were observed in both organs. Thus the relative decrease in granulocyte number was not correlated to a decline in the parasitic load of the fish. To investigate a possible modulatory impact of S. solidus on granulocyte function, head kidney leukocytes were isolated at times before Day 63 p.i. and tested in vitro for their capacity to produce reactive oxygen species (ROS). Head kidney leukocytes from S. solidus-infected fish, analysed immediately after isolation (ex vivo, Day 40 p.i.), exhibited a higher ROS production when stimulated with phorbol myristate acetate (PMA), than leukocytes from naive, sham-treated control fish and fish that had resisted or cleared the infection (exposed but not infected). The latter showed an increased spontaneous ROS production that was not correlated to the numbers of granulocytes present in the head kidney isolates. In infected sticklebacks, spontaneous and PMA-induced ROS production was significantly correlated with numbers of granulocytes present in the head kidney isolates, suggesting that elevated ROS production was due to higher numbers of responding cells rather than an increased capacity of single cells. In vitro, after cultivation for 4 d in the presence of pokeweed mitogen (PWM) or extracts from S. solidus, head kidney leukocytes from control fish showed an increased ROS production and phagocytic activity compared with non-stimulated control cultures. In contrast, head kidney leukocytes from infected fish isolated on Days 48 and 44 p.i., failed to respond to S. solidus antigens in vitro. During S. solidus infection, granulocyte mobilisation resulted in elevated numbers of these cells in head kidneys, but the lack of an in vitro response to S. solidus antigens indicates an in vivo priming of granulocytes by the parasite. These observations may reflect the ability of S. solidus to impair the host's immune response once the parasite is developing in the body cavity of G. aculeatus.     

Transmission of Tetracapsuloides bryosalmonae (Myxozoa: Malacosporea) to Fredericella sultana (Bryozoa: Phylactolaemata) by various fish species

Tetracapsuloides bryosalmonae is a myxozoan parasite of salmonids and freshwater bryozoans, which causes proliferative kidney disease (PKD) in the fish host. To test which fish species are able to transmit T. bryosalmonae to bryozoans, an infection experiment was conducted with 5 PKD-sensitive fish species from different genera. Rainbow trout Oncorhynchus mykiss, brown trout Salmo trutta, brook trout Salvelinus fontinalis, grayling Thymallus thymallus and northern pike Esox lucius were cohabitated with T. bryosalmonae-infected Fredericella sultana colonies and then subsequently cohabitated with statoblast-reared parasite free Bryozoa. Statoblasts from infected colonies were tested by PCR to detect cryptic stages of T. bryosalmonae, which may indicate vertical transmission of the parasite. In this study, brown trout and brook trout were able to infect Bryozoa, while there was no evidence that rainbow trout and grayling were able to do so. Few interstitial kidney stages of the parasite were detected by immunohistochemistry in brown trout and brook trout, while rainbow trout and grayling showed marked proliferation of renal interstitial tissue and macrophages with numerous parasite cells. Intraluminal stages in the kidney tubules were only detected in brown trout and rainbow trout. In contrast to previous observations, pike was not susceptible to PKD in these trials according to the results of T. bryosalmonae-specific PCR. No DNA of T. bryosalmonae was detected in any statoblast.     

Ultrastructure of the penetration of the crayfish integument by the fungal parasite, Aphanomyces astaci, Oomycetes

In the crayfish, Astacus astacus, susceptible to the crayfish plague fungus, penetration of the cuticle by the parasite occurred in the soft cuticle. The zoospore lysed the surface lipid layer, tore it away, and formed an infection peg (germ tube) that penetrated through the epicuticle. A septum was formed in the infection peg, and a hypha was formed below the inner epicuticular surface. In the endocuticle, hyphae grew preferrentially parallel to the surface, occassionally perpendicular to it. Growth direction in relation to cuticle architecture is discussed. Subsequently, some hyphae started to penetrate out through the epicuticle. This process was preceded by the swelling of the hyphal tip touching the inner side of the epicuticle. The hypha penetrating out through the epicuticle was much thicker than the infection peg. Histolytic activity, combined with mechanical penetration, seems to be evident in all stages and levels except in the outward penetration of the epicuticular lipid surface layer, where only mechanical rupture could be seen. Differences in the protoplasmic ultrastructure were found between the spore and the penetrant hyphae. Penetration of the cuticle of a resistant crayfish was essentially identical to that in susceptible ones. However, inward penetration of intact epicuticle was too scarce to allow for ultrastructural studies.     

Cytodiagnosis of eumycotic mycetoma: a case report

BACKGROUND: Mycetoma is a late-stage clinical manifestation of a subcutaneous infection produced by bacteria (actinomycetoma) or fungi (eumycetoma). Only a few articles have described the morphologic appearance of this uncommon pathology on cytology. The distinction between eumycetoma and actinomycetoma in fine needle aspiration cytology (FNAC) is as accurate as in histopathology, as demonstrated in the present case. CASE: A 30-year-old man presented with a large swelling on his left foot with a discharging sinus. FNAC of the swelling yielded pus-like material. Initial Papanicolaou and Giemsa stains showed the presence of septate, branching fungal hyphae and black granules against the inflammatory background. The presence of fungus was confirmed by PAS stain. The species was identified as Exophiala jeanselmei on fungal culture. CONCLUSION: Mycotic mycetoma can be accurately diagnosed by FNAC, which is a simple, inexpensive and rapid technique when there is a high index of suspicion. Special stains and culture studies are helpful in confirmation of diagnosis and species identification.