Birefringent spores differentiate Encephalitozoon and other microsporidia from coccidia

Tissue sections containing protozoa with birefringent spores indicate an infection by microsporidia. Hematoxylin and eosin (HE) does not affect spore birefringence, but some special stains (Goodpasture, Brown and Brenn, or Gram) obscure it. Encephalitozoon cuniculi from an infected puppy, Glugea stephani from the winter flounder Pseudopleuronectes americanus, and Plistophora sp. from the Japanese eel Anguilla japonica all have birefringent spores. Encephalitozoon was studied first and then the two genera from fishes were included for comparison. Small masses of newly formed spores (pseudocysts) line Glugea cysts and then merge into the contents of the cyst as it enlarges and bulges through the intestinal musculature to become subserosal. The birefringence of Plistophora is present in fully mature spores contained in pseudocysts, but may disappear when the spores are released and become involved in granulomas. Coccidians from various hosts were always nonbirefringent. Whenever a protozoan organism in a tissue could be either microsporidian or coccidian, a test for birefringence, if positive, resolves the question. There may be no need to use a special stain.     

Cellular immunity in salmonids infected with the microsporidial parasite Loma salmonae or exposed to non-viable spores

Following a per os challenge of naive rainbow trout with live spores of Loma salmonae, head kidney mononuclear cells (MNC) in culture were able to proliferate in response to crude soluble parasite extract or intact dead spores. A significant response was seen by week 2 post-exposure and a maximum response developed by week 6 or 8, respectively. During this initial challenge, spore filled cysts developed on the gills of challenged fish, and the cysts ruptured by week 12 as is typical for microsporidial gill disease of salmonids (MGDS). Two weeks following this, fish were re-challenged with live spores, and in these fish an enhanced in vitro proliferative response of MNC was immediately apparent, and spore filled cysts did not develop. In contrast, when naive trout were given dead spores by intraperitoneal injection, the most pronounced proliferative responses of MNC developed earlier (week 2 PE) and the response was greater when cells were incubated in vitro with dead spores rather than with crude soluble extract. When these fish were re-challenged per os with live spores, a heightened proliferation in MNC was observed 4 weeks after this exposure and the fish likewise resisted development of xenomas. In fish infected orally or injected intraperitoneally with spores, a marked increase in the response to the mitogen concanavalin A was seen for 22 weeks post-exposure when compared to controls not receiving any spores.     

The effect of dose, route, number of injections and the use of adjuvant on the clearance of and immune response to, haemocyanin following injection into brown trout (Salmo trutta)

The primary and secondary immune responses were investigated in trout injected with haemocyanin (HCN) intramuscularly (IM) or intraperitoneally (IP), with or without adjuvant. HCN was detected in the blood 30 min after injection and clearance times varied after one injection from 8 to 56 days. Fish given two and three injections cleared the antigen faster. Precipitins against HCN were first detected 21 to 30 days after injection and were still present on Day 56. However, antibodies detectable by indirect haemagglutination (IHA) and complement fixation (CFA) were initially demonstrated 7 to 21 days after a single injection and highest titres were reached between 33 and 56 days according to the experimental protocol. In fish given two injections, maximum titres were reached between Days 42 and 56 (IM), and Days 50 and 56 (IP). With three injections, maximum CFA and IHA values occurred on Days 62 and 66 respectively in the case of IP, and on Days 103 and 106 respectively with IM. Overall, higher titres were found with IHA than by CFA.     

Pomegranate enriched diet enhances the hematology, innate immune response, and disease resistance in olive flounder against Philasterides dicentrarchi

Olive flounder, Paralichythys olivaceus fed with pomegranate enriched diet and challenged with or without Philasterides dicentrarchi had a significantly higher white blood cell (WBC) count on weeks 2 and 4 than the infected group fed with non enriched diet (standard diet). Similarly the red blood cell (RBC) counts did not significantly change in control and treated fish on weeks 1 and 2. It was significantly increased in treated fish on week 2 when compared to the control. In both the groups the hemoglobin (Hb) and hematocrit (Ht) levels significantly increased on weeks 2 and 4. The mean corpuscular volume (MCV) did not significantly change at any time in both groups whereas mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) increased significantly on week 4 in the treated group. The leukocytes such as lymphocytes (Lym), monocytes (Mon), neutrophils (Neu), and biochemical parameters such as total protein (TP), glucose (GLU), and calcium (CAL) levels significantly increased in treated groups on week 2 or 4 as compared to the control. The scuticocidal activity and respiratory burst activity were significantly enhanced in treated groups with or without parasite on weeks 2-4. However, the serum lysozyme activity was significantly enhanced from weeks 1 to 4. The protective response in terms of cumulative mortality was low in groups fed with enriched diet against parasite when compared to control. Therefore, we suggest that pomegranate enriched diet following challenge with P. dicentrarchi restores the altered hematological and biochemical parameters, and improves the innate immune system in olive flounder against P. dicentrarchi.     

Development of Tetracapsuloides bryosalmonae (Myxozoa: Malacosporea) in bryozoan hosts (as examined by light microscopy) and quantitation of infective dose to rainbow trout (Oncorhynchus mykiss)

The myxozoan parasite Tetracapsuloides bryosalmonae is the causative agent of proliferative kidney disease (PKD), a highly damaging disease of cultured salmonid fish. Within this study, phylactolaemate bryozoans were collected from a river known to be endemic for PKD and subsequently cultured in the laboratory. Sequential developmental stages of T. bryosalmonae were studied by light microscopy within the living bryozoan colonies, allowing the identification of stages attached to host peritoneum, consistent with previous molecular evidence of cryptic stages. Infection resulted in the production of large numbers of spores, which were released from the bryozoans. Experimental exposure of rainbow trout (Oncorhynchus mykiss) to medium in which infected bryozoans were cultured resulted in clinical PKD. Rainbow trout were exposed to known numbers of T. bryosalmonae spores collected by micromanipulation, which had been released from mature spore sacs within colonies of the bryozoan Fredericella sultana. Exposure to one spore was sufficient to lead to development of PKD. These findings indicate that small numbers of bryozoans are capable of releasing sufficient spores to infect large numbers of fish, having implications for future control methods for PKD in salmonid farming.     

Cardiorespiratory effects and efficacy of morphine sulfate in winter flounder (Pseudopleuronectes americanus)

OBJECTIVE: To assess the cardiorespiratory effects of morphine sulfate and evaluate whether morphine blocks cardiac responses to a noxious stimulus in winter flounder. ANIMALS: 42 winter flounder (Pseudopleuronectes americanus) that were acclimated at 10 degrees C. PROCEDURES: Each fish was fitted with a Doppler flow probe around the ventral aorta; cannulae were placed for injection of drug or saline (0.9% NaCl) solution and assessments of respiration. Selected cardiorespiratory variables were measured in morphine-injected (40 mg/kg, IP [n = 18] or 17 mg/kg, IV [2]) or saline solution-injected (1.6 mL [22]) fish at various intervals. Heart rate and cardiac output (CO) were also measured in flounder that were injected with saline solution (n = 19) or morphine (10) and received a noxious or innocuous stimulus (injection of 5% acetic acid or saline solution SC into a cheek) 50 minutes later. RESULTS: Morphine administration promptly induced marked bradycardia (and a concomitant reduction in CO), followed by prolonged (> 48 hours) increases in CO and heart rate. Morphine injection only transiently affected respiratory rate. Application of a noxious stimulus to control flounder resulted in a significant (10%) but transient (< 5 minutes' duration) increase in CO, which was completely blocked by prior administration of morphine. CONCLUSIONS AND CLINICAL RELEVANCE: Although morphine blocked the response to a noxious stimulus in fish, its cardiovascular effects might preclude its use in many research situations. Investigation of the dose dependency of these cardiovascular effects and their interspecific variation is required to determine the applicability of morphine for use in fish.     

Henneguya corruscans n. sp. (Myxozoa,Myxosporea, Myxobolidae), a parasite of Pseudoplatystoma corruscans (Osteichthyes,Pimelodidae) from the Paraná River,Brazil: A morphological and morphometric study

This paper describes the parasite Henneguya corruscans n. sp. which infects the gills of Pseudoplatystoma corruscans Spix and Agassiz, 1829 found in the Paraná River, Brazil. The parasites belong to the interlamellar-epithelial type as defined by Molnár (2002) [Molnár, K., 2002. Site preference of fish myxosporeans in the gills. Dis. Aquat. Org. 48, 197–207]. The spores examined had thin, smooth walls with symmetric valves; the total length of the spores was 27.6 (25–29) μm. The spore body was ellipsoidal in frontal view and biconvex in lateral view and they measured 14.3 (13–15) μm long by 5 μm wide and 4 μm in thickness. The polar capsules were small and elongated, equally sized, with a rounded posterior extremity and tapering anteriorly, and they corresponded more or less the half the length of the spore body; they were 6.8 (6–7) μm long by 2 μm wide, and the polar filament formed 5–6 coils obliquely to the axis of the polar capsule. The tail was 13.7 (12–15) μm long and bifurcated shortly after the end of the spore body. The importance of the infection for the farming of P. corruscans is discussed.     

Molecular cloning and expression analysis of interferon regulatory factor 8 (IRF8) in turbot, Scophthalmus maximus

Mucus immunoglobulin (Ig) of flounder (Paralichthys olivaceus) was purified by the combination of salting-out, Sephacryl S-300 gel filtration chromatography and DEAE Sepharose chromatography. According to the SDS-PAGE and native-PAGE, the purified mucus Ig showed apparent molecular weights of 72 kDa (heavy chain) and 26 kDa (light chain), and a total molecular weight of 798 kDa, which indicated mucus IgM was in tetrameric form. Purified mucus Ig was used to immunize the Balb/C mice, nineteen hybridomas secreting monoclonal antibodies (mAbs) against flounder mucus Ig were obtained by indirect enzyme-linked immunosorbent assay, and three of them designated as 1A-M2, 1C-M10 and 3F-M9 were cloned by limiting dilution. In Western blotting, the three mAbs specifically reacted to the heavy (H) chain of mucus Ig, but not reacted with serum Ig of flounder, whereas mAb 2D8 against serum Ig previously produced could react with the H chain of both mucus and serum Ig, indicating the composition of the mucus and serum Ig H chains was different. Meanwhile, surface Ig positive (sIg+) lymphocytes in the peripheral blood, spleen, skin and gills of healthy flounder, were analyzed by flow cytometry using mAb 1A-M2 and mAb 2D8, and the results revealed that both mAbs were reactive with the sIg+ lymphocytes. The positive reactivity rates for mAb 1A-M2 were 38.64% in the peripheral blood, 23.6% in the spleen, 16.56% in the skin and 6.26% in the gills, while the positive reactivity rates for mAb 2D8 were 48.89%, 33.7%, 15% and 6.02%, respectively, suggesting mucus Ig was similar, but not identical, to serum Ig. These results generated important mucosal immunological information and gave a valuable insight into understanding the mucosal immunity in flounder.     

Antiparasitic and immunomodulatory effect of innovative treatments against Myxobolus sp. infection in Diplodus puntazzo

The potential antiparasitic and immunomodulatory effect of three treatments against myxosporean parasites on the innate immune system of sharpsnout sea bream (Diplodus puntazzo) was investigated. Fish naturally infected with Myxobolus sp. (Bivalvulida/Platysporina), a histozoic parasite mainly affecting the renal interstitial tissue, were treated by oral administration of a combination of salinomycin with amprolium, Origanum essential oil or fumagillin in a small-scale field trial. Various leucocyte functions influenced by myxosporean infection were examined in order to determine treatment effects on leucocyte immunocompetence of treated fish. One month post treatment all drugs caused a significant decrease in prevalence and intensity of infection in comparison to untreated, infected fish. The effect was most prominent in salinomycin with amprolium treated fish, which 1-month post treatment contained either no cysts at all or a few spores free in melanomacrophage centres revealing almost total elimination of the parasite and the antiparasitic action of the treatment. There was no histopathological evidence of drug toxicity. Antiparasitic action was accompanied by a significant enhancement of phagocytic activity demonstrated by ingestion of large numbers of latex beads and the secretion of high levels of reactive nitrogen intermediates by phagocytes in vitro. Complete restoration of the diminished mitogenic responses and serum lysozyme secretion was also detected in salinomycin with amprolium-treated fish compared to untreated, infected fish. These data suggest that salilomycin with amprolium may be a promising treatment for myxosporean infections in intensively cultured warm-water fish, exhibiting action partially via the enhancement of host, innate immune functions and leading to parasite elimination.     

A Viral Disease in Larvae and Juveniles of the Japanese Flounder Paralichthys olivaceus

Abstract

From 1985 to 1987, outbreaks of a disease resulting in mass mortality occurred in larvae and juveniles of the Japanese flounder Paralichthys olivaceus cultured at prefectural and private hatcheries in Hiroshima Prefecture, Japan. The disease occurred in 10-30-d-old fish that were reared at about 18–20°C, and mortality usually reached 80–90% in a few weeks. The affected fish had opaque fins and a hyperplastic epidermis on the fins and skin. Electron microscopy revealed hexagonal virus particles in the nucleus (100–140 nm in diameter without an envelope) and cytoplasm (190–230 nm with an envelope) of the affected epidermal cells. Although isolation of the causative agent by the use of five fish-cell cultures was not successful, the disease was transmitted to healthy larval flounder by exposing them to a 0.45-μm filtrate of diseased fish homogenate. The agent's morphological features and its sensitivity to ether, to a pH of 3, and to a 30min exposure to 50°C indicate it is a herpesvirus.     

粉纹夜蛾培养细胞对家蚕微孢子虫的吞噬及与孢壁蛋白的关系

解明家蚕微孢子虫(Nosema bombycis,Nb)入侵宿主细胞的相关因素,有助于深入研究Nb对家蚕的侵染机制。利用激光共聚焦显微镜观察Nb孢子接种粉纹夜蛾(Trichoplusiani,Tn)培养细胞48h后孢子的分布状态和培养细胞的生长情况,并对接种Nb孢子后的Tn培养细胞培养物总蛋白进行SDS-PAGE分析,证实经0.2mol/LKOH预处理的Nb孢子可以被Tn培养细胞所吞噬。对Nb孢子孢壁蛋白的SDS-PAGE分析显示,Nb孢子经KOH预处理后,大小为80kD和12kD的孢壁蛋白带缺失或明显减弱,30kD的孢壁蛋白带丰度降低。推测KOH预处理使Nb孢子部分孢壁蛋白被分解(部分或完全)或使蛋白结构发生变化,由此影响了Nb孢子与Tn培养细胞的相互作用关系,有利于Tn培养细胞对Nb孢子的吞噬。     

Detection of Myxobolus (Myxosoma) cerebralis in Salmonid Fishes in Oregon

Abstract

Myxobolus (Myxosoma) cerebralis, the etiological agent of whirling disease, was detected in salmonid fish populations in northeastern Oregon. This is the first record of M. cerebralis in the Pacific Northwest of the USA. During an epizootiological survey for the parasite, two methods for spore detection were compared, and an efficient procedure for determining M. cerebralis infection in adult fish was developed. The enzyme digest method was more efficient than the plankton centrifuge procedure for examination of numerous individual lots of fish processed during the survey. Sampling only the area around the otoliths was at least as effective as sampling entire heads for detection of spores in infected fish.     

Surveillance for Ceratomyxa shasta in the Puget Sound watershed, Washington

Discovery of fish exhibiting clinical signs of ceratomyxosis in Washington State prompted concern over the potential impact of the myxozoan parasite Ceratomyxa shasta on native stocks of steelhead Oncorhynchus mykiss (anadromous rainbow trout). To investigate these concerns, a survey of 16 freshwater systems within the Puget Sound watershed, including Lake Washington, was conducted by sentinel exposure of susceptible fish (cutthroat trout O. clarkii and rainbow trout). Fish were exposed for 7 d during September 2003 and May 2004 and then were returned to a holding facility for monitoring of disease signs. Mortality caused by the parasite occurred only in the exposure group held at the University of Washington Hatchery, which receives its water from Portage Bay of Lake Washington. Fish from all other sites were negative for C. shasta, both visually and by polymerase chain reaction (PCR) assay, except for a single fish held at the Tumwater Falls Hatchery in September 2003. A single deformed spore was detected in that fish, but infection could not be confirmed by PCR and the parasite was not detected from any other fish held at that site during either the September or the May exposure. From these results, we conclude that C. shasta is not likely to have contributed significantly to the decline of steelhead populations throughout Puget Sound.     

Production of Ceratonova shasta Myxospores from Salmon Carcasses: Carcass Removal Is Not a Viable Management Option

Severe infection by the endemic myxozoan parasite, Ceratonova (synonym, Ceratomyxa) shasta, has been associated with declines in and impaired recovery efforts of populations of fall-run Chinook Salmon Oncorhynchus tshawytscha in the Klamath River, California. The parasite has a complex life cycle involving a polychaete worm host as well as a salmon host. Myxospore transmission of this parasite, from salmon to polychaete, is a life cycle step during which there is a potential for applied disease management. A 3-year data set on prevalence, intensity, and spore characteristics of C. shasta myxospores was obtained from adult Chinook Salmon carcasses surveyed in the main stem of the Klamath River and three of its tributaries, Bogus Creek and the Shasta and Trinity rivers. Annual prevalence of myxospore detection in salmon intestines ranged from 22% to 52%, and spore concentration values per intestinal scraping ranged from 3.94 × 10² to 1.47 × 10⁷ spores. A prevalence of 7.3% of all carcasses examined produced >5.0 × 10⁵ spores, and these carcasses with “high” spore counts accounted for 76–95% of the total spores in a given spawning season. Molecular analysis of visually negative carcasses showed that 45–87% of these samples had parasite DNA, indicating they contained either low spore numbers or presporogonic stages of the parasite. Myxospores were rarely found in carcasses of freshly spawned adults but were common in decomposed carcasses of both sexes. The date of collection or age (based indirectly on FL) did not influence detection. The longer prespawn residence time for spring-run Chinook Salmon compared with that for fall-run Chinook Salmon in the Trinity River was associated with higher spore loads. The dye exclusion method for assessing spore viability in fresh smears indicated an inverse relationship in spore integrity and initial spore concentration. A carcass-removal pilot project in Bogus Creek for 6 weeks in the fall of 2008 (907 carcasses removed) and 2009 (1,799 carcasses removed) failed to measurably influence the DNA quantity of C. shasta in targeted waters. Combined with the high numbers of carcasses that contributed myxospores, we therefore deemed that this labor-intensive approach is not a viable management option to reduce the infectivity of C. shasta in Chinook Salmon in the Klamath River.

Received January 23, 2015; accepted September 28, 2015     

Experimentally Induced Whirling Disease I. Dose Response of Fry and Adults of Rainbow Trout Exposed to the Triactinomyxon Stage of Myxobolus cerebralis

Abstract

The intensity and prevalence of whirling disease was tested by exposure of 2-monthold fry and 1-, 2-, and 3.5-year-old adults of rainbow trout Oncorhynchus mykiss to a known number of laboratory-produced Myxobolus cerebralis at the actinosporean triactinomyxon stage. Fry exposed to graded concentrations of infectivity (triactinomyxons) for 3 h were individually examined for spores of Myxobolus cerebralis 5 and 6 months later. Exposure of fish to the lowest doses, 1 and 10 triactinomyxons per fish, did not result in detectable myxosporean spores. Fish that became lightly infected by a dose of 100 triactinomyxons per fish experienced a decrease in the incidence of infection between 5 and 6 months after exposure. A linear relationship was found between the numbers of recovered myxosporean spores and doses of 100–10,000 triactinomyxons per fish, and the spore burden appeared to plateau at doses of 10,000–100,000 triactinomyxons per fish. Adult fish continuously exposed to the highest dose of triactinomyxons for 3.5 months were infected and asymptomatic, however, the severity of myxosporean infection decreased with increased age of fish. This information may help in controlling whirling disease in salmonids.     

Evaluation of nested polymerase chain reaction, pepsin/trypsin digest, and histology for the detection of Myxobolus cerebralis in salmonid fishes of Indiana and Michigan.

The objectives of this study were to survey fish from state hatcheries in Indiana and Michigan and to compare the nested polymerase chain reaction (PCR) test with pepsin/trypsin digest (PTD) and histopathology for the diagnosis of whirling disease (WD). One group of 40 and 9 groups of 60 fish heads, for a total of 580 samples, were submitted from hatcheries in Indiana and Michigan. These samples were examined for myxozoan spores using histopathology, PTD, and PCR tests. The heads were hemisectioned, and one half was fixed in 10% neutral-buffered formalin for histopathologic examination. The other half was processed for PTD. Some of the sediment was examined for the presence of myxozoan spores, and the rest was prepared for the nested PCR. Histologic examinations did not reveal Myxobolus cerebralis in any of the 580 samples. One hundred serial step sections, taken at 5-microm intervals, were evaluated for samples with positive spore identification by PTD. Histologic examination of these sections failed to reveal any myxozoan parasites. Myxozoan spores were observed in 16.9% (98/580) of samples in sediment after PTD. Spores morphologically similar to those of M. cerebralis were observed in 1.0% of PTD samples (n = 6). The nested PCR indicated that M. cerebralis spores were present in 0.5% of samples (n = 3). All 3 nested PCR-positive samples came from the same hatchery, however, spores of M. cerebralis were seen in 1 sample, spores of other myxozoan species were seen in the second sample, and spores were not seen in the third sample. When comparing the PTD to the nested PCR test, the PTD diagnosed 1 true positive, 5 false positives, 2 false negatives, and 572 true negatives, for a sensitivity of 33% and a specificity of 99.1%. Screening for M. cerebrallis infection in this study indicated a low prevalence of the disease. Histopathology was a very insensitive indicator of WD. The PCR test was highly specific and was used to differentiate spores of M. cerebralis from similar spores of other species.     

The effect of growth hormone-releasing peptide-2 (KP102) administration on plasma insulin-like growth factor (IGF)-1 and IGF-binding proteins in Holstein steers on different planes of nutrition

This study was conducted to investigate the nutrition-dependent changes in insulin-like growth factor (IGF)-1 and IGF-binding proteins (IGFBPs) with growth hormone releasing peptide-2 (D-Ala-D-betaNal-Ala-Trp-D-Phe-Lys-NH(2); GHRP-2 or KP102) treatment in growing Holstein steers. Eight 13 month-old Holstein steers were grouped on two levels of feed intake (high intake (HI); 2.43% body weight or low intake (LI); 1.22%) and each group was daily injected with KP102 (12.5 microg/kg body weight/day) or saline solution into the jugular vein during 6-day period. The concentration of plasma GH showed an increase after an i.v. bolus injection of KP102 on Day 1 and Day 6 in both the LI and HI groups. Plasma IGF-1 began to increase 10 hr following an i.v. bolus injection of KP102, but this was only observed in the HI group (P < 0.05). Also, the plasma IGF-1 in the HI group with daily injections was significantly greater than the LI group from Day 1 of KP102 administration (P < 0.05). It reached maximum values of 125.1 +/- 7.6 ng/ml after Day 2, and returned to pre-injection levels after Day 4, however, no change in plasma IGF-1 was observed in LI with administration of KP102. During 6 days of treatment, plasma 38-43 kDa IGFBP-3 and 24 kDa IGFBP-4 were significantly higher in KP102 treated steers but only in the HI group (P < 0.05). Plasma 34 kDa IGFBP-2 decreased in the HI group and did not show any change following an injection of KP102. In conclusion, the effect of stimulated endogenous GH with KP102 administration increased plasma IGF-1, 38-43 kDa IGFBP-3 and 24 kDa IGFBP-4 levels in the HI group of growing Holstein steers, but not in the LI one. Thus, we strongly believe that the plasma IGF-1 and IGFBPs response to KP102 treatment is modulated by the nutritional status of growing Holstein steers and the increased plasma IGF-1 concentration with KP102 treatment may be regulated by plasma 38-43 kDa IGFBP-3 and 24 kDa IGFBP-4 in Holstein steers.     

The effect of porcine somatotropin supplementation in pigs on the lipid profile of subcutaneous and intermuscular adipose tissue and longissimus muscle.

The effect of porcine somatotropin (pST) on the lipid profiles of adipose tissue and muscle was investigated. Sixteen crossbred barrows were injected daily with either 3 mg of pST or a placebo. After slaughter, total lipid and fatty acid composition of raw subcutaneous (SC) adipose and intermuscular (IM) adipose tissue and longissimus muscle were determined. The SC adipose tissue from pST-treated pigs had a 7.5% decrease in total lipid content; specific fatty acids 16:0, 18:0, and 18:1(n-9)c decreased most. The IM fat from pST-treated pigs had lower levels of 16:0 and 20:0. There was no effect of pST treatment on the lipid profile of the longissimus muscle. The data suggest that pST treatment produces small but significant changes in the saturated fatty acid content of adipose tissue in pigs.     

The seasonal antibody response in juvenile summer flounder (Paralichthys dentatus) to the hemoflagellate Trypanoplasma bullocki

An immuno-blot assay was used to investigate the serum antibody response in flounder injected with formalin-killed flagellates (immunized) and those injected with saline (control) and challenged with live T. bullocki after 21 days. Fish were held at 20 degrees C and at ambient temperature from October through June. At 20 degrees C immunized fish had significantly higher antibody titers than control fish, but immunized fish were not protected from infection with T. bullocki. At ambient temperature, after initial flagellate growth phase, antibody titer varied directly with temperature (2-25 degrees C) and T. bullocki intensity varied inversely with titer. Flagellates were eliminated from the peripheral circulation in both immunized and control fish when antibody titer peaked in May. Recovered fish were immune to homologous challenge for at least one year.