Corynebacterium suis infection in swine. 1. Clinical diagnosis with special consideration of urine studies and cystoscopy

Experimental and spontaneous infections with Corynebacterium suis in sows were investigated. In early stages animals show no clinical disorders or only for a short time. However, there are already marked changes in urinary samples (hematuria, proteinuria, leukocyturia, gross alterations). Using an endoscope mucosal irritations can be seen mainly on the floor of the bladders. In chronic cases alterations in urine are more pronounced. If a pyelonephritis is present in addition to the cystitis, general signs of illness are evident including anorexia, emaciation, anemia, subnormal body temperature and abortions. Bladders demonstrate an erosive and ulcerative, hemorrhagic cystitis on the whole mucosal surface. Uremia appears only in late stages of the disease.     

Endoscopic studies of the bladder in breeding sows--diagnosis of cystitis

With a flexible scope the examination of the bladder is possible at the standing sow without anaesthesia. When using a non-flexible scope general anaesthesia with a supplementary spinal anaesthesia is necessary in order to avoid lesions of the urinary tract and damage of the instrument. For a systematic inspection the bladder must be emptied and filled with air. The state of the bladder can be estimated by the colour of mucosa, the condition of visible blood vessels and of the mucosal surface. To prevent infections careful disinfection of the scope as well as antibiotic treatment of the bladder is important. There were good correlations between endoscopic findings in sows with cystitis and parameters of urinalysis especially for sensory parameters, proteinuria, leukocyturia and significant bacteriuria. An advantage of cystoscopy is the possibility to survey beginning or chronic symptoms of cystitis, even when urine is nearly unchanged.     

Mass mortality associated with a Sphaerospora-like myxosporidean infestation in juvenile cobia, Rachycentron canadum (L.), marine cage cultured in Taiwan

Cultured cobia, Rachycentron canadum , of 45–80 g exhibited anaemia and ascites, and a mottled red and grey, extremely enlarged kidney with cream-coloured patches or spherical nodules. Cumulative mortality was about 90% within 1 month. Extrasporogonic or sporogonic stages of a myxosporean appeared in the blood, glomerulus, renal tubules and renal interstitium. The renal tubules were the main target tissue of the parasite and were completely occluded by sporogonic pseudoplasmodia at various degrees of maturity. Many sporogonic stages were attached to the brush border of the epithelium of the renal tubules. Mature spores were seen in the lumen of the tubules. They were elongated or spherical with numerous refractile granules in the cytoplasm. The polar filament formed 3–5 coils. No bacteria or viruses were isolated from the diseased fish. Based on the results of microbiological, histopathological and electron microscopical examinations, the cobia disease was believed to be caused by a Sphaerospora -like myxosporean. This is the first report of a myxosporean in cobia in aquaculture.     

雄性羊驼尿生殖道和阴茎的形态特征

为探索羊驼易发尿道阻塞而致膀胱破裂的形态学特征,采用大体解剖学和普通石蜡切片技术,对雄性羊驼的尿生殖道和阴茎进行了研究,并与反刍动物的相关结构进行了比较。结果表明,雄性羊驼的尿生殖道分骨盆部和阴茎部,阴茎部靠近皮下,且背侧与骨性骨盆联合紧贴,外伤和出血均易导致阻塞;阴茎在阴囊后形成乙状弯曲。组织学发现尿生殖道骨盆部黏膜为变移上皮,阴茎部海绵体和阴茎海绵体结构相似,阴茎头末端皮下有一层软骨组织。     

An immune-complex glomerulonephritis of Chinook salmon, Oncorhynchus tshawytscha (Walbaum)

Chinook salmon from New Zealand were shown to have a generalized membranous glomerulonephritis that was most severe in large fish. Marked thickening of the glomerular basement membrane was the most consistent lesion, with the presence of an electron-dense deposit beneath the capillary endothelium.Severely affected glomeruli also had expansion of the mesangium and loss of capillaries,synechiae of the visceral and parietal epithelium and mild fibrosis of Bowmans capsule. Chinook salmon from British Columbia, Canada with bacterial kidney disease caused by Renibacterium salmoninarum had similar histological lesions. They also had thickened glomerular basement membranes that were recognized by rabbit antiserum to rainbow trout immunoglobulin. This was true only when frozen sections of kidney were used and not formalin-fixed tissue. An attempt to experimentally produce a glomerulopathy in rainbow trout by repeated immunization with killed R. salmoninarum was not successful. Case records from the Fish Pathology Laboratory at the University of Guelph over a 10-year period revealed that a range of species were diagnosed with glomerulopathies similar to those seen in Chinook salmon. The majority of these cases were determined to have chronic inflammatory disease. This report has identified the presence of immunoglobulin within thickened basement membranes of Chinook salmon with glomerulonephritis and supports the existence of type III hypersensitivity in fish.     

Gross, histological and scanning electron microscopic appearance of dorsal fin rot in farmed Atlantic salmon, Salmo salar L., parr

The gross, histological and scanning electron microscopic appearance of dorsal fin rot in farmed Atlantic salmon parr, Salmo salar L. is described. The lesions were grouped into seven categories: (1) peripheral erosion and ray splitting; (2) peripheral erosion with some nodularity; (3) severe nodularity with differing degrees of tissue loss; (4) extensive to total loss of the dorsal fin; (5) smooth thickening of the dorsal fin; (6) haemorrhagic dorsal fin lesions; and (7) healed dorsal fin rot lesions. The main sign of injury was clefts extending through the epithelium. These injuries were consistent with bites from other parr. During healing from such wounds, damaged cells sloughed from the surface, and there was swelling and hyperplasia in the remaining cells. The majority of the thickening in the fins was the result of epithelial hyperplasia with a variable cellular inflammatory response. The distal epithelium of fins with severe fin rot (i.e. nodular and eroded) was rough, irregular and swollen with superficial nodular extensions. Wounds in all stages of repair were more numerous in such areas. Fin rays were frequently observed protruding from the abnormal epithelium at the distal edge of the fin. With the exception of the isolation of Aeromonas salmonicida from a small number of cases, no significant bacterial involvement was detected. Under scanning electron microscopy, bacteria were only detected on the exposed fin rays and not in association with the abnormal epithelium. In the majority of cases, the dorsal fin was either the only fin damaged or the most severely damaged. It is suggested that the hyperplastic response to numerous bite wounds is responsible for the accumulation of abnormal epithelium typical of dorsal fin rot in farmed Atlantic salmon parr.     

大黄鱼(Pseudosciaena crocea)内脏白点病的组织病理和超微病理分析

利用组织切片及超薄切片电镜技术对患内脏白点病大黄鱼(Pseudosciaena crocea)的肝脏、脾脏和肾脏3种组织进行病理学分析,探讨该病的致病机理.结果显示,大黄鱼的临床症状为体表无明显病症,脾、肾、肝等内脏有大量白色结节;组织病理显示,肝、肾和脾是感染损伤的主要靶器官,出现组织变性坏死,空泡化严重,炎性细胞浸润;病变组织均出现病理性结节.超微病理显示,病鱼肝、肾、脾细胞超微结构受损严重,尤以线粒体和细胞核损伤明显.线粒体肿胀,嵴断裂消失,空泡化;细胞核核膜破裂,染色质浓缩边集;肾脏和脾脏均发现大量菌聚集成团的病原.研究表明,大黄鱼内脏白点病的组织细胞病理变化特征显示了病原菌的入侵和危害,造成鱼体生理代谢紊乱,免疫力和抵抗力下降,鱼体终因无法维持正常的生命活动而导致死亡.     

卵形鲳鲹刺激隐核虫病组织病理学研究

利用组织切片、HE染色和显微观察的方法,研究了卵形鲳鲹患刺激隐核虫病组织病理变化。组织切片结果可见,感染鱼的鳃和体表受到直接机械损伤;鱼体在缺氧状态下,入侵的刺激隐核虫有可能对机体产生某种毒素,也可能因刺激隐核虫感染而引发细菌或病毒继发性感染,造成肝、脾和肾脏不同程度的间接性损伤。表现为鳃部机能严重受阻,肝脏脂肪肝变性严重,肝索紊乱,血窦缩小或消失;脾脏组织结构不清晰,有充血、出血和坏死等症状;肾脏结构不清晰,肾小管及肾小体缩小,肾小管细胞融合(黏连),组织间隙松散扩大,局部细胞坏死,存在脂肪粒。     

Myxosporean infection causing intestinal disease in farmed turbot, Scophthalmus maximus (L.), (Teleostei: Scophthalmidae)

The present report describes an intestinal disease which causes important losses in farmed turbot. Mortality rates were higher in summer and reached 100% in all tanks where the disease was confirmed. Affected fish showed external signs consisting of anorexia, sunken eyes and a typical prominent bony ridge on the skull. These signs can be considered the pathognomonic signs of the disease, together with the gut lesions seen in the histological study. Pallor of the internal organs, intestinal haemorrhages and the presence of liquid in the intestine were also observed, with ascites in heavily infected fish. Histopathological damage was evident in the gut, with severe enteritis, detachment of epithelium, haemorrhages and inflammation of the subepithelial connective tissue. The myxosporean aetiology was demonstrated in all the fish showing the characteristic signs of disease. Myxosporean stages, including scarce spores, were found in the affected epithelium or free in the intestinal lumen together with epithelial debris. The present study demonstrates the importance of this myxosporean disease which represents a serious threat for turbot culture. This is also the first record of a member of the Myxosporea in turbot.     

Pathogenesis of vibriosis in cultured juvenile red abalone, Haliotis rufescens Swainson

Abstract. Morbidity of intensively cultured red abalone, Haliotis rufescens , as well as experimentally stressed (elevated temperature and hyper-oxygenation) abalone, was studied using clinical, histological, immunofluorescent and bacteriological techniques. Histological study showed a typical pattern of bacterial infection from all groups studied, characterized by epithelial exfoliation or rupture and systemic growth of the bacteria along vascular sinuses and along neural sheaths. Peripheral neurons degenerated rapidly and a responsive host cellular infiltrate did not appear to effectively retard the advancement of the infection. Nine bacterial isolates from the culture system water or sick animals were characterized biochemically. All were Gram-negative, facultatively anaerobic, non-aerogenic, oxidase-positive rods with single polar flagella and thus appeared related to the Vibrio group. Further characterization showed that most isolates did not correspond to specifically characterized vibrios, Antiserum prepared to the isolates contained antibody both to common group antigens (from all nine strains) and to strain-specific antigens. Selection of antiserum and subsequent absorption permitted the use of the antiserum for specific recognition of each isolate. Immunofluorescent studies clearly demonstrated that antiserum to an isolate corresponding to Vibrio alginolyticus was the predominant antiserum producing positive staining of infecting bacteria in the typical lesions in abalone tissues. The pattern of positive staining corresponded to histopathological observations of the disease. The disease can be managed in husbandry systems by both limiting the number of potentially pathogenic bacteria and by limiting the exposure of the animals to physico-chemical stresses.     

Study of the distribution of active caspase‐3‐positive cells in turbot,Scophthalmus maximus (L.), enteromyxosis

Enteromyxosis caused by Enteromyxum scophthalmi is one of the parasitizations with a higher economic impact on turbot, Scophthalmus maximus (L.), aquaculture. This myxosporean produces severe catarrhal enteritis with abundant inflammatory infiltrates in the lamina propria‐submucosa (LP), epithelial detachment and leucocyte depletion of the lymphohaematopoietic organs. Some advances made on the pathogenesis pointed to a role of apoptosis in the enteromyxosis. Therefore, the main aim of this work was to employ the TUNEL assay and the anti‐(active caspase‐3) immunohistochemical assay to detect apoptotic cells in both healthy and E. scophthalmi‐infected turbot in order to establish the presence and distribution of apoptotic cells during development of the disease. More apoptotic cells located within the gastrointestinal epithelium were observed in the initial stages of the infection in E. scophthalmi‐infected turbot compared with non‐infected turbot. As the infection progressed, a higher degree of apoptosis occurred in the epithelium of folds heavily parasitized. In the severely infected turbot, apoptosis was also found among the leucocytes of the intestinal inflammatory infiltrates. Moreover, the number of active caspase‐3‐positive cells in the lymphohaematopoietic organs tended to increase with disease severity. In view of the results, increased apoptosis in the epithelium may favour the scaling that occurs during enteromyxosis and cell death of leucocytes in the intestinal LP, contributing to leucocyte depletion in severe cases.     

Initial swim bladder inflation in the larvae of Tilapia mossambica (Peters) and Morone saxatilis (Walbaum)

Tilapia and striped bass larvae inflate their swim bladders on the 7th–9th and 5th–7th days after hatching, respectively. The primordial bladder of Tilapia has no pneumatic duct and larvae do not gulp atmospheric gas for the initial swim bladder inflation. Prominent columnar epithelium is characteristics of the bladder of Tilapia prior to inflation. Hypoxic conditions inhibit inflation and result in degeneration of the columnar epithelium and the irreversible malfunction of the swim bladder. The swim bladder of the striped bass has a glandular epithelium and a pneumatic duct prior to inflation. The mode of the initial inflation and functional role of the pneumatic duct remain unclear, although strong aeration and turbulent water in the rearing containers were observed to be main factors enhancing normal inflation of the larval swim bladder.     

酸性磷酸酶在文昌鱼体内的分布

文昌鱼代表由无脊椎动物进化到脊椎动物的过渡类型。关于文昌鱼体内酸性磷酸酶(ACP)的分布至今未见报道。本实验采用酶组织化学方法,首次报道ACP在文昌鱼体内的分布。结果表明,文昌鱼表皮、消化道(包括肠和肝盲囊)、生殖腺和神经系统中具有极强的ACP活性,内柱细胞顶部具有较强的ACP活性,围鳃腔上皮和鳃上皮细胞中也具有一定的ACP活性。文昌鱼体内ACP的主要功能可能与细胞内消化和免疫活动有关。     

Pathogenicity of Pasteurella sp. in lumpsuckers (Cyclopterus lumpus L.)

The incidence of disease caused by Pasteurella sp. in farmed lumpsuckers in Norway has been steadily increasing in recent years, causing significant economic losses and fish welfare issues. The disease affects all life stages, both in hatcheries and after release into salmon cages. Therefore, it is important to establish robust challenge models, to be used for vaccine development. Exposure experiments via intramuscular and intraperitoneal injection underlined the high virulence of the bacteria, whereas the cohabitation and bath models allowed the chronic symptoms of the disease to be studied more accurately. Skin lesions and haemorrhage at the base of fins were observed in the more acute cases of the disease. Symptoms including white spots over the skin, especially around the eyes, characterized the chronic cases. The latter were most prominent from the bath challenge model. Histopathology indicated a systemic pattern of disease, whereas qPCR analysis from head kidney showed that bacteria may be present in survivor fish at the end of the challenges. In all the challenge models investigated, Pasteurella sp. was re‐isolated from the fish, thus fulfilling Koch's postulates. These findings highlight the importance of screening of lumpsuckers prior to transfer to minimize the risks of carrying over asymptomatic carriers.     

Pathology of bacterial gill disease: sequential development of lesions during natural outbreaks of disease

Abstract. This study examines the sequential pattern of bacterial colonization and the sequential development of branchial lesions during typical outbreaks of bacterial gill disease (BGD) among four groups of commercially reared rainbow trout fingerlings, Oncorhynchus mykiss (Richardson). During a 5-month monitoring regime, prior to the onset of natural outbreaks of BGD, gill morphology of examined fish remained unaltered. Bacterial colonization of the gills was immediately preceded by several gill changes which were widely present in fish from all groups under study. These changes, which were detectable at the ultrastructural level only, included cytoplasmic blistering and degeneration of the microridges of the superficial filament epithelium, in addition to slight topographical irregularity of the filament tips suggestive of mild hyperplasia. Bacterial colonization began at these altered filament tips before spreading to more proximal regions of the filament and adjacent lamellar surfaces. Explosive increases in proportional morbidity and mortality coincided with the development of the following gill lesions: extensive bacterial colonization of lamellar surfaces, lamellar epithelial hydropic degeneration and necrosis, and lamellar oedema. In contrast, gill lesions such as lamellar fusion, epithelial hyperplasia and various metaplastic responses were detected later either as subacute (3–5 days) or chronic (7–14 days) changes. The role of epithelial necrosis as an important event during BGD is discussed as a possible mechanism leading to the development of sub-acute and chronic gill lesions.     

Vibriosis in channel catfish, Ictalurus punctatus (Rafinesque)

Abstract. An epizootic due to Vibrio anguillarum was observed in channel catfish, Ictalurus punctatus (Rafinesque). Lesions in infected fish included ulceration and petechiae on body surfaces, vent and caudal peduncle. Internally, haemorrhages in liver and kidney were present and the intestinal tract was filled with a clear viscous fluid. Using an isolant recovered during the epizootic, laboratory studies were conducted to determine (a) bacterial dynamics of the agent in blood, kidney and liver and (b) clinical haematologic and biochemical parameters in infected fish. Under conditions of the study, vibrios were apparently sequestered in kidney and liver during initial stages of infection (8–12 h after exposure). Later, bacterial numbers in blood were comparable to those in kidney and liver. Clinical parameters of infected catfish were suggestive of cellular and tissue destruction and renal dysfunction. Based upon data of the present study and those of others, vibriosis appears to be a disease in which the agent is localized in select tissues. Secondary septicaemia may be incidental to factors which compromise host defences.     

Changes in the intestinal bacterial community during the growth of white shrimp,Litopenaeus vannamei

In this study, we documented the changes in the intestinal bacterial community at four stages in Litopenaeus vannamei: 14 days postlarvae (L14) and 1‐, 2‐ and 3‐month old juveniles (J1, J2, J3), using 454 pyrosequencing techniques. The intestinal bacterial community was dominated by three bacterial phyla, Proteobacteria, Bacteroidetes and Actinobacteria at all stages. However, the relative abundance and bacterial lineages varied at the family level. The intestinal bacterial community of L14 and J1 was similar, with dominant members belonging to the Comamonadaceae of Betaproteobacteria. Conversely, bacterial members affiliated to Flavobacteriaceae of Bacteroidetes were dominant in J2 and Vibrionaceae of Gammaproteobacteria was dominant in J3. The abundance of Microbacteriaceae of Actinobacteria also fluctuated during the four stages. Bacterial members of Flavobacteriaceae and Rhodobacteraceae (Alphaproteobacteria) were present through all growth stages, and likely form the intestinal core microbiome of L. vannamei. However, they varied at the operational taxonomic unit (OTU) level through the growth stages. The intestinal bacterial community of pond‐rearing shrimp included the three main bacterial phyla identified above, and an additional group, Mycoplasmataceae of Mollicutes. Our results demonstrate that the intestinal bacterial community of L. vannamei was highly dynamic during the growth stages. Bacterial members belonging to Commamonadaceae dominated in the earlier growth stage of shrimp, possibly influenced by feeding with Artemia nauplii, but there was a shift to Flavobacteriaceae in the mid and Vibrionaceae in the late growth stages.     

Morphological characterization of the adherence and invasion of Streptococcus agalactiae to the intestinal mucosa of tilapia Oreochromis sp.: An in vitro model

Streptococcosis in tilapia Oreochromis sp. is possibly the most important bacterial disease for fish production worldwide. In Colombia, streptococcosis is caused by Streptococcus agalactiae (GBS), but in other countries, Streptococcus iniae is also involved. Prevention of streptococcosis is required and must be addressed for economic, social, international trade and public health reasons. This research used an in vitro culture of tilapia intestine to detail the intestinal mucosal response once the pathogen contacts the epithelium. We show that S. agalactiae sheds off its capsule to adhere to the epithelium. The bacterium adheres as a single individuum, in groups or in chains and is able to divide on the apical border of enterocytes. GBS adheres at and invades exclusively through the apical portion of the intestinal folds, using the transepithelial route. Once within the cytoplasm of enterocytes, the bacteria continue to divide. On the basolateral side of the epithelium, the microorganisms leave the cells to reach the propria and travel through the microcirculation. No evidence of an immuno‐inflammatory reaction or goblet cell response in the epithelium or the lamina propria was seen during the process of adherence and invasion of the pathogen.     

鲤慢性喹乙醇中毒的病理学和组织残留

试验以含10、25、50、100、150和200mg·kg-1喹乙醇的饲料对鲤进行了慢性毒性试验。经90d的试验,各组的发病率分别为17.5%、27.5%、35.0%、42.5%、47.5%和55.0%,死亡率分别为5.0%、10.0%、12.5%、17.5%、20%和30.0%。中毒鱼表现为特征性的"应激性出血综合征",且Hb含量和RBC数量降低,血清AST、ALT活性升高。病理组织学表现为鳃小片水肿,上皮细胞增生、变性、坏死和脱落。心肌纤维、肾小管上皮细胞和肾上腺细胞空泡变性;肝细胞脂肪变性或水泡变性及溶解坏死;肠道发生卡他性肠炎;超微结构上,心肌纤维、肝细胞和肾小管上皮细胞的线粒体肿胀、嵴断裂、溶解,粗面内质网扩张,核糖体脱颗粒;肝细胞内脂滴明显增多;肠上皮微绒毛脱落、崩解和肠上皮细胞大量坏死脱落。中毒鱼肝、肾和肌肉组织内喹乙醇含量与对照组之间有显著性差异。     

Improved disease resistance in Rana rugulosa (Daudin) after β-glucan administration

There is an increasing problem with bacterial disease associated with stress in farmed frogs. In general, little is known about the immune system in farmed tropical ranid species and, in particular, about its functional ability. As part of a study on the non-specific immune system, immunostimulation by β-glucan was evaluated in preventing bacterial disease in cultured ranid frogs. Fifty frogs were grown to a mean weight of 40 g and separated into three experimental groups. These were exposed to a motile aeromonad bacterial challenge alone (Au), bacterial challenge with β-glucan (Au + glucan) or saline only (controls) to serve as controls. There were four replicate tanks containing five frogs per tank in each experimental group. After bacterial challenge, all the animals were observed twice daily, and the number of mortalities per tank was recorded. The liver was sampled in order to recover bacteria, and any colonies grown were identified. The survival of the β-glucan-treated frogs was significantly higher than that of the bacterial challenge group alone.