Development of a whole-body cortisol extraction procedure for determination of stress in golden shiners, <Emphasis Type="Italic">Notemigonus crysoleucas</Emphasis>

Baitfish such as golden shiners are subjected to stress during harvesting, grading, and transport. Their small size makes it difficult to measure the stress response with the biological indicator cortisol using conventional assay methods for plasma. This paper examines the development and validation of methods for whole-body cortisol extraction from individual baitfish. Three types of extracts were tested: (1) an ethyl ether unaltered extract (UA); (2) an extract reconstituted in phosphate buffered saline (PBS); (3) an extract that had been increased in volume by the addition of food-grade vegetable oil (VO). These extracts were evaluated using validation tests with radioimmunoassays (RIA) and enzyme-linked immunosorbent assays (ELISA). The UA extract produced inadequate volumes of extract for multiple assays and could not be used for the determination of cortisol in a single fish. The PBS reconstitution method failed the precision recovery of serial dilutions (62.3%), linearity (R ²: 0.7864), and parallelism validation tests. The VO volume-boosting method passed all validation tests [intra-assay coefficent of variation (%CV): 16.3 for ELISA and 5.9 for RIA; inter-assay %CV: 10.3; spiked recovery: 102.0%; dilution recovery: 93.0%; linearity R ²: 0.9435; log of serial dilutions was parallel] and provided enough extract for multiple assays from an individual baitfish. Based on these results, we conclude that the VO volume-boosting method presents a means for determining cortisol from individual baitfish using either RIA or ELISA assays.     

Validation of a whole-body cortisol extraction procedure for channel catfish (<Emphasis Type="Italic">Ictalurus punctatus</Emphasis>) fry

We validated a whole-body cortisol extraction technique for channel catfish, Ictalurus punctatus, fry. Three volume enhancement methods were tested: CAL method (zero calibrator A diluent added to lipid extract), PBS method (phosphate buffered saline added to lipid extract), and VO method (food grade vegetable oil added to lipid extract). The volume enhancement extracts were evaluated using a commercial radioimmunoassay kit. Sensitivity, accuracy, precision, reproducibility, and parallelism could not be determined for the PBS method as cortisol levels were not detected in any of the extracted samples. Intra-assay coefficient of variation (CV) for the CAL and VO methods were 7.3 and 8.3%, respectively, while inter-assay CV were 9.6 and 10.6%, respectively. Based on the sensitivity, accuracy, precision, reproducibility, and parallelism results, we conclude that the CAL method is the most appropriate method for volume enhancement of catfish fry lipid extract. Using the CAL method to detect cortisol in catfish fry, fish were stressed daily for 2 weeks. Fry weights were similar throughout the study while whole-body cortisol levels were higher (P < 0.01) in stressed fish after 1 day of stress. These data show the CAL method can effectively measure whole-body cortisol in catfish fry.     

Alternative matrices for cortisol measurement in fish

Plasma cortisol is the most commonly used indicator of stress in fish but, as the blood sampling procedure itself can be a source of stress, it would be helpful to measure cortisol using less invasive matrices. It is also necessary to find alternative matrices as stress indicators in dead fish in which blood sampling is impossible. In the present study, we investigated transport stress in three aquaculture species, European sea bass (Dicentrarchus labrax L.), common carp (Cyprinus carpio L.) and rainbow trout (Oncorhynchus mykiss Walbaum), by cortisol determination (radioimmunoassay) in plasma and other matrices (skin mucus, gut content, lateral muscle and caudal fin). Cortisol significantly increased after transport in all species and matrices, except in the sea bass gut content, where it remained unchanged. The three species responded to transport stress by producing different cortisol levels. In conclusion, the significant correlation found between plasma cortisol and most of the other matrices opens up the possibility of using them to evaluate stress in fish: mucus sampling is a less invasive method than blood sampling, and in addition to muscle and fin sampling, it can be used in postmortem fish.     

Effect of plant density in coupled aquaponics on the welfare status of African catfish,Clarias gariepinus

We investigated the effects of plant density on the welfare of African catfish, Clarias gariepinus, in coupled aquaponics over 85 days. The moderate density (mpd) of basil, Ocimum basilicum, was compared with the high density (hpd) and control (n = 0). The behavior was analyzed by visual and video observations, and after the application of induced stressors, skin injuries, blood glucose, lactate, and plasma cortisol responses were considered. The hpd fish showed the least activity (control: visual 77.8%, video 81.6%; mpd: 74.6%, 82.6%; hpd: 63.2% [p < 0.05], 78.8%). High agonistic behavior (control: 5, 131; mpd: 4, 57; hpd: 1, 45) and the highest number of injuries (control: 3.9; mpd: 2.9; hpd: 3.4) were observed in the control. Glucose and lactate levels did not differ significantly (control: 5.5, 2.6 mmol/L; mpd: 5.6, 2.7 mmol/L; hpd: 5.3, 2.6 mmol/L); however, cortisol levels did (control: 18.8 ng/mL, mpd: 19.9 ng/mL, hpd: 25.8 ng/mL). pH adjustment led to additional stress, resulting in temporal cortisol alterations. While in the control and mpd, low cortisol levels were followed by acute responses and downregulation, the hpd fish showed prior elevation and lagged an acute response. However, comparing injuries and behavioral patterns with control, aquaponics with high basil density influenced African catfish positively.     

Evaluation of the INNO‐LiPA mycobacteria v2 assay for identification of aquatic mycobacteria

Fifty‐seven isolates of mycobacteria comprising 10 reference strains, 47 field isolates and one non‐Mycobacterium isolate were screened using commercial INNO‐LiPA v2 assay kits. All mycobacteria isolates tested hybridized with the Mycobacterium genus probe on the LiPA strip. All M. marinum, M. fortuitum and M. chelonae reference and field strains and three out of the four M. gordonae isolates hybridized to their corresponding species‐ or complex‐specific probes. Two cultures (a type strain and a field isolate) yielded mixed growth of two mycobacterial species, i.e. M. chelonae and M. fortuitum. A Mycobacterium isolate from one of these cultures was subsequently purified and correctly identified with the kit. However, sequence analysis of the 16S–23S rRNA internal transcribed spacer (ITS) region of various mycobacteria isolates revealed a misidentification of M. shottsii and M. pseudoshottsii with the kit because these isolates reacted with the M. marinum/M. ulcerans probe. Moreover, nine of the 13 field isolates presumed to be M. fortuitum from the results of the kit had closer ITS sequence homology with M. conceptionense, a species which, to our knowledge, has never been reported in fish. These findings highlight the need to redesign the M. fortuitum–M. peregrinum probe included in the INNO‐LiPA assay and to introduce additional complex‐specific probes into the kit. Nevertheless, the kit proved to be a rapid and reliable method for identifying mycobacteria in the aquatic environment and would be particularly useful in laboratories without sequencing facilities.     

Immunological and physiological validation of enzyme-linked immunosorbent assay for growth hormone of the Asian catfish, Clarias batrachus

This study describes the development and validation of competitive antigen capture enzyme-linked immunosorbent assay (ELISA) for growth hormone (GH) of the catfish, Clarias batrachus. Isolated GH was characterized first through bioassay, amino acid sequencing, immunoblotting and immunocytochemistry, and then used to raise antibody to develop ELISA. The lowest detection limit of the assay system was 0.17 ng ml⁻¹, and the standard curve had an ED₅₀ value of 0.35 ng ml⁻¹. Repeated determination of GH in a plasma pool exhibited intra- and interassay co-efficient of variation of 8.1% and 7.09% (n=5), respectively. Dose–response inhibition curves resulting from dilutions of plasma and pituitary homogenate of C. batrachus were parallel to the standard curve, while such parallelism in the case of Cyprinus carpio and H. fossilis was absent revealing no cross reaction of them in the present ELISA. In vivo effects of sGnRH, testosterone, 5-HT and morphine on plasma GH were also examined to validate the ELISA physiologically. sGnRH, testosterone and morphine increased, while 5-HT decreased GH level in a dose-dependent manner.     

Mycobacteria in aquarium fish: results of a 3‐year survey indicate caution required in handling pet‐shop fish

Fish are commonly infected with non‐tuberculous mycobacteria (NTM), which should be regarded as potential pathogens when handling aquarium fish and equipment. This study examined 107 aquarium fish from pet shops. Cultivation of the fish samples using different selective media was conducted for identification of NTM. Isolates were identified using the GenoType Mycobacterium common mycobacteria and additional species assays, sequencing of the 16S rRNA and rpoB genes, and real‐time PCR assay for identification of Mycobacterium (M.) marinum. Among the investigated fish, 79.4% (85/107) were positive for mycobacteria, with 8.2% (7 of 85) having two mycobacterial species present. Among the positive fish, the common pathogens M. marinum, Mycobacterium fortuitum (M. fortuitum group) and Mycobacterium chelonae were identified in approx. 90% of fish and other NTM species in 10%, including Mycobacterium peregrinum/septicum, Mycobacterium gordonae, Mycobacterium arupense, Mycobacterium kansasii, Mycobacterium ulcerans and Mycobacterium setense. The well‐known human pathogen M. marinum was present in 10.6% of the positive fish (9 of 85). The species of mycobacteria identified in the study are not only recognized as aquarium fish pathogens, but can also cause pathology in humans. Microbiological and clinical communities should therefore be sensitized to the role of NTM in infections associated with exposure to aquarium fish.     

Validation of diagnostic assays to screen broodstock for Flavobacterium psychrophilum infections

It is hypothesized that the frequency of bacterial coldwater disease outbreaks can be reduced through the detection of the aetiologic agent, Flavobacterium psychrophilum, in broodstock followed by culling of eggs from heavily infected broodstock. Before a culling programme can be instituted, however, it is necessary to determine the sensitivity and specificity of existing assays for the detection of F. psychrophilum. In this study, tissue and ovarian fluid samples were collected from 224 fish at five hatcheries and screened using an enzyme‐linked immunosorbent assay (ELISA), a membrane‐filtration fluorescent antibody test (MF‐FAT), bacteriological culture and nested PCR. Latent class analysis was used to estimate sensitivity and specificity of kidney culture, kidney ELISA, nested PCR and MF‐FAT. Analytical sensitivity of the ELISA varied but was greatest when bacteria were cultured under iron‐limiting conditions. Diagnostic sensitivity estimates ranged from 0.02 (kidney culture) to 0.97 (kidney ELISA). Specificity estimates ranged from 0.02 (MF‐FAT) to 0.98 (kidney ELISA). In a separate challenge experiment, the ELISA confirmed the presence of F. psychrophilum in sub‐clinically infected fish. Results from this study demonstrate that the ELISA is an appropriate tool to screen broodstock and provides an indication of infection severity, which is crucial for implementation of a screening/culling programme.     

Prothrombin time, activated partial thromboplastin time and fibrinogen values in Mediterranean marine teleosts

Coagulation profiles for marine fish were determined with the standard global screening assays, the prothrombin time (PT) and activated partial thromboplastin time (APTT) using homologous tissue thromboplastin and phospholipid extracts to investigate species-specificity of brain thromboplastins, and to establish a base-line coagulation profile for marine fish of economical importance for Mediterranean aquaculture (sea bream, Sparus aurata, sea bass, Dicentrarchus labrax, red porgy, Pagrus pagrus, common dentex, Dentex dentex). Sea bass and sea bream displayed the shortest PT (9.0–15.0 s) and APTT (22.1–35.8 s) in comparison to the other species (PT: 24.8–37.2 s, APTT: 39.6–66.4 s). Red porgy displayed the lowest fibrinogen value (1.2–2.3 g l^–1), while no differences were found among the other species (2.2–2.8 g l^–1). It is concluded that the interaction of the tissue factor and plasma clotting factor is relatively species specific and that the use of homologous brain extracts is preferable for haemostasis study in marine fish.     

Estimating Amino Acid Requirement of Brazilian Freshwater Fish from Muscle Amino Acid Profile

Information on nutritional requirement of some Brazilian farmed fish species, especially essential amino acids (EAA) requirements, is scarce. The estimation of amino acids requirements based on amino acid composition of fish is a fast and reliable alternative. Matrinxa, Brycon amazonicus, and curimbata, Prochilodus lineatus, are two important Brazilian fish with potential for aquaculture. The objective of the present study was to estimate amino acid requirements of these species and analyze similarities among amino acid composition of different fish species by cluster analysis. To estimate amino acid requirement, the following formula was used: amino acid requirement = [(amount of an individual amino acid in fish muscle tissue) × (average totalEAA requirement among channel catfish, Ictalurus punctatus, Nile tilapia, Oreochromis niloticus, and common carp, Cyprinus carpio)]/(average fish muscle totalEAA). Most values found lie within the range of requirements determined for other omnivorous fish species, in exception of leucine requirement estimated for both species, and arginine requirement estimated for matrinxa alone. Rather than writing off the need for regular dose–response assays under the ideal protein concept to determine EAA requirements of curimbata and matrinxa, results set solid base for the study of tropical species dietary amino acids requirements.     

Humoral immune response of Japanese eel, Anguilla japonica Temminck & Schlegel, to Pleistophora anguillarum Hoshina, 1951 (Microspora)

A humoral response of Japanese eel, Anguilla japonica Temminck & Schlegel, to the microsporean Pleistophora anguillarum Hoshina was demonstrated using immunoblotting and an enzyme-linked immunosorbent assay (ELISA). Japanese eel immunoglobulin was purified by affinity chromatography. The immunoglobulin was composed of 25-kDa light chains and 72-kDa heavy chains. The ELISA values of P. anguillarum antibodies in naturally infected fish sera were significantly higher than those of clinically healthy fish. Spore proteins from the microsporean were separated by electrophoresis and subjected to analysis by Western blot. Sera from naturally infected fish showed different reaction patterns to the spore proteins. While the sera randomly selected from naturally infected eels all showed a significant positive reaction to P. anguillarum antigens, the mucus from only three out of the nine infected eels reacted positively in the ELISA test. Subsequent analyses indicated that there was no significant difference in the amount of mucus immunoglobulin among the tested eels. Therefore, the generally lower ELISA values of mucosal anti-P. anguillarum antibodies from the infected eels tested were evidently not caused by a lack of immunoglobulin per se, but seem to be the result of a lack of anti-P. anguillarum antibodies in the mucus and/or a lower affinity in the anti-P. anguillarum antibodies that were present.     

Bench‐top validation testing of selected immunological and molecular Renibacterium salmoninarum diagnostic assays by comparison with quantitative bacteriological culture

No gold standard assay exhibiting error‐free classification of results has been identified for detection of Renibacterium salmoninarum, the causative agent of salmonid bacterial kidney disease. Validation of diagnostic assays for R. salmoninarum has been hindered by its unique characteristics and biology, and difficulties in locating suitable populations of reference test animals. Infection status of fish in test populations is often unknown, and it is commonly assumed that the assay yielding the most positive results has the highest diagnostic accuracy, without consideration of misclassification of results. In this research, quantification of R. salmoninarum in samples by bacteriological culture provided a standardized measure of viable bacteria to evaluate analytical performance characteristics (sensitivity, specificity and repeatability) of non‐culture assays in three matrices (phosphate‐buffered saline, ovarian fluid and kidney tissue). Non‐culture assays included polyclonal enzyme‐linked immunosorbent assay (ELISA), direct smear fluorescent antibody technique (FAT), membrane‐filtration FAT, nested polymerase chain reaction (nested PCR) and three real‐time quantitative PCR assays. Injection challenge of specific pathogen‐free Chinook salmon, Oncorhynchus tshawytscha (Walbaum), with R. salmoninarum was used to estimate diagnostic sensitivity and specificity. Results did not identify a single assay demonstrating the highest analytical and diagnostic performance characteristics, but revealed strengths and weaknesses of each test.     

Ammonia‐, Sodium Chloride‐, and Calcium Sulfate‐induced Changes in the Stress Responses of Jundiá, Rhamdia quelen,Juveniles

Salt (NaCl) and gypsum (CaSO₄) are used as water additives to mitigate fish stress and improve specimen survival. High stocking densities and the transportation of fish can increase aqueous ammonia, which can, in turn, alter fish cortisol secretion. The objectives of this study were to assess the effects of salt, gypsum, and aqueous ammonia on some stress‐induced physiological responses of jundiá, Rhamdia quelen, juveniles induced by captivity and handling, and to determine the lethal ammonia concentration for this species. Jundiá juveniles were subjected to the following five treatments: water only, water + ammonia (0.4 mg/L), water + NH₃ + NaCl (6 g/L), water + NH₃ + gypsum (150 mg/L), and water + NH₃ + NaCl + gypsum. Blood samples were taken after intervals of 1, 5, 24, and 96 h, and the concentrations of cortisol, glucose, chloride, ammonia, and hematocrit were determined. The NH₃ LC₅₀ value after 96 h of exposure (LC_50?96h) was measured to be 1.9 mg/L NH₃. Either salt or gypsum reduced both cortisol and glucose levels during most of the experimental period, but the combination of both reduced these levels even further. The combined use of NaCl and CaSO₄ demonstrates a synergic effect on mitigating stress responses induced by handling and aqueous ammonia in jundiá juveniles.     

基于DNA条形码的黑龙江中游漂流性鱼卵鉴定及其群落结构初探

黑龙江位于高纬度寒冷地区, 鱼类自然繁殖及早期资源发生具有其独特性, 了解漂流性鱼卵种类及群落结构的空间格局, 对于保护黑龙江鱼类资源具有重要意义。于 2022 年 6—7 月对黑龙江干流的抚远、勤得利、萝北 3 个江段开展漂流性鱼卵资源监测研究, 采用 DNA 条形码技术鉴定鱼卵的种类, 获得 CO I 基因序列 1179 条, 共鉴定鱼类 15 种, 隶属于 2 目 3 科 8 亚科 15 属, 其中鲤科最多(13 种), 占 83.29%; 鳅科和鮨科各 1 种, 分别占 16.45% 和 0.25%。分析结果显示, 漂流性鱼卵的种类组成主要以贝氏䱗(Hemiculter bleekeri)、花斑副沙鳅(Parabotia fasciatus)、潘氏鳅鮀(Gobiobotia pappenheimi)等小型鱼类为主, 群落多样性为抚远江段最高, 抚远以上呈下降趋势, 表现出明显的空间异质性。鲢(Hypophthalmichthys molitrix)、草鱼(Ctenopharyngodon idella)、鳡(Elopichthys bambusa)、翘嘴鲌(Culter alburnus)、鳊(Parabramis pekinensis)、鳜(Siniperca chuatsi)等重要经济鱼类的鱼卵相对重要性指数(IRI)由抚远(下游)至萝北(上游)呈显著下降趋势, 暗示抚远江段的繁殖亲鱼的规模要大于以上江段, 而萝北江段资源面临衰退。本研究推测, 环境因子及过度捕捞应该是影响重要经济鱼类早期资源的主要因素, 建议在产卵场设立禁渔区、禁渔期, 降低捕捞强度, 开展鱼卵、仔稚鱼等早期资源的长期监测研究, 以科学保护并合理利用黑龙江渔业资源。     

Age determination and growth of Pacific bluefin tuna, Thunnus orientalis, off Japan and Taiwan

Age determination of wild captured Pacific bluefin tuna, Thunnus orientalis, was conducted using sagittal otoliths of 806 specimens (47–260 cm in fork length) caught in the waters off Japan and Taiwan. Otoliths were transversely sectioned and the opaque and translucent zones were analyzed. Opaque zones mainly appeared on the otolith edge from April to July, indicating that the opaque zone is formed annually. The opaque zones formed during later life (age 10+) were more distinct than the earlier zones. The estimated ages of specimens ranged from 1 to 26 years. Parameters of the von Bertalanffy growth function were estimated to be 249.6 cm, 0.173, and −0.254 years for L_∞, k, and t₀, respectively. Growth of younger fish was rapid up to 5 years old attaining about 150 cm, and then growth rate decreased. After that, fish attained about 200 cm at 9 years old and about 225 cm (90% of L_∞) at 13 years old (50% of maximum age). This paper updates the biological information on length at age with a large size range to support stock assessment model analyses for this commercially valuable species.     

Morphology and distribution of granulomatous inflammation in freshwater ornamental fish infected with mycobacteria

Mycobacteriosis in fish is a chronic progressive ubiquitous disease caused by Mycobacterium marinum, M. gordonae and M. fortuitum in most cases. The aim of this study was to describe the morphology and distribution of lesions in 322 freshwater ornamental fish across 36 species. Granulomatous inflammation was diagnosed by gross examination and histopathology testing in 188 fish (58.4%); acid‐fast rods (AFR) were determined in only 96 (51.1%) fish from 19 species after Ziehl–Neelsen staining. The most often affected organs with AFR were the kidney (81.2%), digestive tract (54.1%), liver (48.2%), spleen (45.9%) and skin (21.2%); sporadically, AFR were found in the branchiae (9.4%) and gonads (4.7%). In 14 randomly selected fish originating from four different fish tanks, the distribution of mycobacterial infection was studied by culture examination of the skin, gills, muscle tissue, digestive tract, liver, spleen and kidney. In 12 fish, the species M. marinum, M. gordonae, M. fortuitum, M. triviale, and M. avium subsp. hominissuis (serotypes 6 and 8 and genotype IS901? and IS1245+) were detected; mixed infection caused by different mycobacterial species was documented in five of them.     

Prevalence of nematode parasite,Contracaecum, in Nile tilapia,African catfish and Barbus species in Lake Hawassa,Ethiopia

A cross‐sectional study was conducted from November 2017 to March 2018 at Lake Hawassa to determine the prevalence of larval Contracaecum infestation in Nile tilapia, (Oreochromis niloticus), African Catfish (Clarias gariepinus) and Barbus species (Barbus intermedius). Fifty‐two point six per cent (52.6%) previous prevalence and 5% precision were used to estimate the sample size. Accordingly, a total of 383 randomly sampled fish species comprising of 163 (42.6%) C. gariepinus, 159 (41.5%) O. niloticus and 61(15.9%) B. intermedius were examined. The length and weight of each sampled fish were recorded. All the sampled fish were examined for the evidence of Contracaecum parasites in Hawassa University Veterinary Parasitology and Pathology Laboratory. Chi‐square values and comparison of proportions were used to analyse the data. The overall prevalence of Contracaecum parasites of fish population was 31.6%. The distribution of parasite was significantly affected by fish species (p = .000), sexes (p = .018), length (p = .003) and weight classes of fish (p = .026). As a hygienic problem and gutting activity conducted at the sides of the Lake Hawassa and distribution of discarded fish wastes for surrounding piscivorous birds by butchers and other people, the life cycle of Contracaecum parasite was perpetuated. Thus, the parasite is of zoonotic significance. Therefore, awareness creation activities for societies and control of fish parasites should be conducted in the study area.     

Effects of butaphosphan and cyanocobalamin combination on plasma immune and biochemical parameters of Olive flounder (Paralichthys olivaceus) subjected to crowding stress

The study evaluated the effects of intramuscular injection of butaphosphan and cyanocobalamin combination (BPC) on the plasma levels of cortisol, cytokines, and the activities of selected enzymes in Oliveflounder exposed to crowding stress. Three groups of fish (n = 12 per group) were kept in different glass tanks with stocking densities of 46 kg/m³(group BP and SCH) and 15 kg/m³ (group SCL). Group BP was treated with 0.5 ml of BPC. While groupSCH and SCL were treated with 0.5 ml of saline. Blood was collected a day after injection (T1), and weekly for two consecutive weeks (T2 and T3). The plasma levels of interleukin (IL) ‐1β, IL‐6, cortisol, lysozyme, and the activities of aspartate aminotransferase (AST) and alanine transaminase (ALT)were determined using an ELISA kit. The statistical significance of differences was assessed using ANOVAfollowed by Tukey‐HSD test (P < 0.05 considered significant).A significantbody weight gain was measured in the BP and SCL groupscompared with SCH at T3. The activities of AST (at T1, T2, and T3) and ALT (at T1 and T2) were declined significantly (P < 0.05) following treatment of overcrowded fish with BPC than saline. Similarly, BPCcaused a significant reduction in the levels of cortisol and IL‐1β at T2 and T3.This is the first report of the effect of the combination of butaphosphan and cyanocobalamin on plasma immune and biochemical parametersin fish. Therefore, the combination could be beneficial in preventing overcrowding‐induced immune suppression and tissue damage in Olive flounder.     

Development of an Indirect ELISA to Detect Humoral Response to Flavobacterium columnare Infection of Channel Catfish,Ictalurus punctatus

ABSTRACT

The humoral antibody response of channel catfish, Ictalurus punctatus, to experimental Flavobacterium columnare infection was measured in control (non-infected) and infected (intraperitoneal- and intramuscular-injected) fish by an indirect enzyme-linked immunoassay (ELISA). The antibody response of the experimentally infected fish was significantly higher (P<0.0001) than that of the non-infected channel catfish by both indirect ELISA and agglutination. The indirect ELISA utilized goat anti-channel catfish IgM and a commercially available rabbit anti-goat IgG enzyme conjugate. The antibody response was directed against a cell-free sonicated antigen preparation derived from live whole F. columnare cells. Indirect ELISA and agglutination results correlated (r² = 0.60) for anti-F. columnare antibody response in experimentally and non-infected fish. We were also able to correlate ELISA and agglutination results of 60 fish naturally infected with F. columnare(r² = 0.76). Indirect ELISA will allow for rapid monitoring of humoral antibody, following natural exposure or vaccination against F. columnare, with a small sample of serum.     

Fish hosts of the freshwater mussel Unio foucauldianus Pallary, 1936

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