Neobenedenia melleni from reef ornamental fish species in a retailer of Southeastern Brazil and its possible role as a mechanical vector of bacterial infection.

Annually, more than 2500 ornamental fish species are traded worldwide. Forty percent of these are from marine water. Some 98% of marine species are wild-caught from their natural habitat, and the majority subsequently exported. Wild fish frequently carry pathogens, which could induce diseases after the stress of capture. Neobenedenia melleni is a platyhelminth that mainly attaches to the skin and eyes of the host. It provokes dermal inflammation, epidermal loss, skin depigmentation, reduction in the number of mucous cells, and, consequently, decreased mucus protection, and declining immunological barriers. This makes fish susceptible to secondary infections. A total of 47 wild reef fish from a retailer were examined, suspected to be infected with ectoparasites. The morphological identification revealed N. melleni as a monogenean agent. One monogenean specimen was collected from the eye of each of the 40 fish analyzed to evaluate possible bacterial secondary infections using the matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) technique. The MALDI-TOF MS identified that 59% of monogenean collected from the eyes had bacteria, including some pathogenic to fish. This led us to believe that the ectoparasite can be a possible mechanical vector of pathogenic bacteria for fish culture and maintenance. The use of praziquantel as an antiparasitic agent is also discussed.

Corrigendum: Short and Long-Term Effects of Anesthesia in Octopus maya (Cephalopoda, Octopodidae) Juveniles.

[This corrects the article DOI: 10.3389/fphys.2020.00697.].

Short and Long-Term Effects of Anesthesia in Octopus maya (Cephalopoda, Octopodidae) Juveniles.

This study aimed to explore different substances (or cold sea water) as potential anesthetic agents to facilitate short-term handling in Octopus maya juveniles. We investigated oxygen consumption before (baseline), during (first 600 s of exposure) and after anesthesia (recovery) of octopuses (n = 98; 1.67 ± 0.5 g) exposed to cold sea water (SW; 11 and 13°C), ethanol (EtOH; 0.5; 1.5 and 3.0%), magnesium chloride (MgCl2; 0.75; 1.5 and 3.75%), ethanol combined with magnesium chloride (Mix; 1.5:0.75%; 0.75:1.13%; and 2.25:0.37%) and clove oil (0.15 mL L-1). After exposure, the animals were handled for 180 s (exposed to air) and weighted. Two experimental groups not exposed to anesthetics (with or without handling) were also evaluated. The criteria for general anesthesia were analysed. Times of induction and recovery, incidence of attack response after recovery and possible longer-term effects of repeated general anesthesia on growth and mortality of the octopuses were evaluated. During anesthesia, O. maya juveniles exposed to SW (11 and 13°C), EtOH (0.5; 1.5 and 3.0%), Mix (0.75:1.13%), and clove oil, presented a significant decrease on oxygen consumption. In animals exposed to different concentrations of EtOH and Mix 0.75:1.13%, this decrease was registered after an increase on oxygen consumption. Animals exposed to MgCl2 did not show significant changes on oxygen consumption, except for animals exposed MgCl2 3.75%, which showed a significant increase on oxygen consumption. At the end of recovery, except for octopuses exposed to clove oil and MgCl2 0.75%, the values of oxygen consumption observed were comparable to the ones registered during baseline. Animals exposed to SW 11°C, EtOH 3.0%, Mix 1.5:0.75% and MgCl2 3.75% fulfilled the criteria defined for general anesthesia. Exposure to MgCl2 (all concentrations), SW 13°C and clove oil reduced or inhibited the incidence of attack response after recovery. Except for animals exposed to clove oil, growth of the juveniles was not affected by the exposure to the different substances. Short-term handling (180 s) of O. maya juveniles can eventually be carried out without anesthesia. However, to facilitated handling, we suggest the use of EtOH 3.0% or cold sea water 11°C.

The Chelating Mineral on Organic Acid Salts Modulates the Dynamics and Richness of the Intestinal Microbiota of a Silver Catfish Rhamdia quelen.

The aim of this study was to evaluate the influence of the chelating mineral on propionic acid, calcium or sodium on the composition, dynamics and richness of the intestinal microbiota of a native silver catfish Rhamdia quelen through high-throughput sequencing (HTS). A total of 225 fish (8.43 ± 0.18 g) were distributed in tanks, 15 fish per tank in five groups with three replicates each: Control, Ca-propionate 0.25% (Ca0.25%) Ca-propionate 1% (Ca1%), Na-propionate 0.25% (Na0.25%) and Na-propionate 1% (Na1%). The feed was provided four times a day for 60 days. After experimental period, the fish were fasted for 24 h and the intestine was aseptically collected, pooled by treatment, and fixed in pure absolute ethanol for subsequent DNA extraction and HTS. The HTS showed that the supplementation of the propionic acid chelated to the mineral calcium or sodium in the different concentrations increased the operational taxonomic units and richness in comparison to control group. The main phyla found were Fusobacteria, Firmicutes, Proteobacteria and Bacteroides. Both the fusobacteria and the genus Cetobacterium, especially C. somerae, were positively modulated with Ca0.25% and Na1% supplementation. It can be emphasized that supplementation with calcium or sodium propionate at different concentrations changed the natural microbiota of R. quelen.

Dermocystidium sp. in the gills of farmed Oreochromis niloticus in Brazil.

The genus Dermocystidium is very comprehensive in the host and site of infection, however this is the first report of the occurrence of Dermocystidium sp. in the gills of Nile tilapia. This study was carried out in a fish farming located in the state of Santa Catarina, Brazil. No mortalities were reported in the facility studied and the animals were clinically healthy. During the histopathological analysis of the gills, 8.33% of the fish presented spores of Dermocystidium sp. in the gill tissue. The spores reported herein had a mean length and width of 6.206 x 5.233 µm and a refractile body diameter of 1.965 µm and were studied by histopathology and Transmission Electron Microscopy. This study highlights the importance of a new branchial pathogen in farmed tilapia, as well as to its pathogenic potential, considering the outbreaks of mortalities associated with other fish species.

Three previous recorded species of Dactylogyrus Diesing, 1850 (Monogenea: Dactylogyridae) infecting cultured Carassius auratus in southern Brazil.

The present study aimed to contribute to the monogenean dactylogyrids that occur in C. auratus from an ornamental fish farm in southern Brazil. Samples of goldfish were obtained from a fish farm and the gills were removed and analysed to identify the species of Dactylogyrus to determine the levels of infestation. Three species of Dactylogyrus were found in the gills: Dactylogyrus anchoratus, Dactylogyrus baueri and Dactylogyrus formosus. The most prevalent species was D. baueri [prevalence (P) = 70%, mean intensity (MI) = 2.14 ± 1.21, mean abundance (MA) = 1.5 ± 1.43], followed by D. anchoratus (P = 60%, MI = 1.67 ± 0.81, MA = 1.00 ± 1.05) and D. formosus (P = 30%, MI = 1.66 ± 1.15, MA = 0.5 ± 1.64). Dactylogyrus anchoratus shares morphological characteristics with D. arcuatus by the similarity in shape of the haptor parts. Dactylogyrus baueri shows similarity to D. dulkeiti in relation to the pair of anchors and accessory piece of the male copulatory complex. Dactylogyrus formosus presents an accessory piece with branch and total length of the anchors smaller than D. anchoratus. This study reports the occurrence of D. anchoratus, D. baueri and D. formosus in goldfish cultured in southern Brazil. These monogenean species have been described in cyprinid fishes that have a wide geographic distribution with the result of the dissemination of the host in the world.

A longitudinal assessment of host-microbe-parasite interactions resolves the zebrafish gut microbiome's link to Pseudocapillaria tomentosa infection and pathology.

BACKGROUND: Helminth parasites represent a significant threat to the health of human and animal populations, and there is a growing need for tools to treat, diagnose, and prevent these infections. Recent work has turned to the gut microbiome as a utilitarian agent in this regard; components of the microbiome may interact with parasites to influence their success in the gut, meaning that the microbiome may encode new anthelmintic drugs. Moreover, parasite infections may restructure the microbiome's composition in consistent ways, implying that the microbiome may be useful for diagnosing infection. The innovation of these utilities requires foundational knowledge about how parasitic infection, as well as its ultimate success in the gut and impact on the host, relates to the gut microbiome. In particular, we currently possess limited insight into how the microbiome, host pathology, and parasite burden covary during infection. Identifying interactions between these parameters may uncover novel putative methods of disrupting parasite success. RESULTS: To identify interactions between parasite success and the microbiome, we quantified longitudinal associations between an intestinal helminth of zebrafish, Pseudocapillaria tomentosa, and the gut microbiome in 210 4-month-old 5D line zebrafish. Parasite burden and parasite-associated pathology varied in severity throughout the experiment in parasite-exposed fish, with intestinal pathologic changes becoming severe at late time points. Parasite exposure, burden, and intestinal lesions were correlated with gut microbial diversity. Robust generalized linear regression identified several individual taxa whose abundance predicted parasite burden, suggesting that gut microbiota may influence P. tomentosa success. Numerous associations between taxon abundance, burden, and gut pathologic changes were also observed, indicating that the magnitude of microbiome disruption during infection varies with infection severity. Finally, a random forest classifier accurately predicted a fish's exposure to the parasite based on the abundance of gut phylotypes, which underscores the potential for using the gut microbiome to diagnose intestinal parasite infection. CONCLUSIONS: These experiments demonstrate that P. tomentosa infection disrupts zebrafish gut microbiome composition and identifies potential interactions between the gut microbiota and parasite success. The microbiome may also provide a diagnostic that would enable non-destructive passive sampling for P. tomentosa and other intestinal pathogens in zebrafish facilities.

Trace elements and microbiological parameters in farmed Nile tilapia with emphasis on muscle, water, sediment and feed.

Fish may bioaccumulate contaminants from the aquatic environment and extend them to the food chain provoking risks to human health. This study evaluated the microbiological parameters of the pond´s water and trace elements concentrations in samples of water, sediment, feed and muscle of farmed Nile tilapia used for human consumption in southern Brazil. A total of 240 fish were collected from 12 tilapia farms. Sediment, tank water and dry ration used in the animals' diet were collected for analysis. Analysis were performed by Energy Dispersion X-ray Fluorescence (EDXRF), Inductively Coupled Plasma Mass Spectrometer (ICP-MS), Induced Coupled Plasma Optical Emission Spectrometer (ICP-OES), and Atomic Absorption Spectrometer (AAS-VGA). In addition, the microbiological analysis of the water was carried out. The concentrations of Se, I, Fe, Cu and Zn in fish muscle were higher than the recommended by the Brazilian legislation, considering the advised daily intake for adults. The arsenic element had concentrations above the limit stipulated by the present Brazilian legislation, observed in all samples of muscle, sediment and tank water highlighting a possible environmental and fish contamination by the toxic element. Moreover, the arsenic concentration in the water presented a positive correlation (ρ = 0.33) with arsenic in the muscle, suggesting that tilapia is a good environmental bioindicator, once they properly reflect the levels of arsenic in the water. It is suggested to perform an arsenic speciation for quantification of the inorganic form and accurate assessment of the degree of toxicity in the muscle samples and risks it can bring to human health. Regarding the other potentially toxic elements (Hg, Pb and Cd), and microbiological analysis of water it was verified that the consumption of the fish in question does not raise risks, since the values are within a quality benchmark established by law. The concentration of total and fecal coliforms in pond´s water in the facilities was in agreement with the microbiological indexes required by the legislation of CONAMA class II. Western region presented the lowest concentrations of fecal coliforms when compared to the other regions. There was no significant difference in the microbiological counts of total heterotrophic bacteria, Vibrio spp. and Pseudomonas spp. among the regions.

Hematology and condition factor of tui chub and fathead minnow parasitized by nematode from Upper Klamath Lake, Oregon, USA.

This study evaluated the hematological profile and condition factor (Kn) of tui chub Siphateles bicolor and fathead minnow Pimephales promelas and their associations with larvae of Contracaecum sp. infection of the heart. A total of 30 tui chub and 17 fathead minnow were collected from Upper Klamath Lake, Oregon, USA, measured, and weighed and blood was drawn for hematological analysis. Nematode larvae parasitized tui chub with a prevalence of 50% and mean intensity of 1.40, while 11.8% of fathead minnow were parasitized at a mean intensity of 1.0. Non-parasitized tui chub were significantly larger than the parasitized fish, indicating that small fish could be easily predated by the definitive host, a piscivorous bird. Although the relatively large worm occupied a large portion of the atrium, the presence of the larvae did not affect tui chub Kn, possibly associated with low parasite intensity and a harmonic co-evolution. Only parasitized fathead minnow showed significant differences in red blood cell measurements (greater cell width and larger nuclei) compared to non-parasitized fish. Lymphocytes were the most common white blood cells found in tui chub, followed by neutrophils, monocytes, and periodic acid-Schiff positive granular leukocytes; in fathead minnow lymphocytes were followed by heterophils, monocytes, neutrophils and eosinophils. This study is the first report of Kn and description of blood cells and hematological parameters in these fish species.

Rocinela signata (Isopoda: Aegidae) parasitizing the gills of the spotted goatfish Pseudupeneus maculatus (Actinopterygii: Mullidae) in Northeastern Brazil.

Parasitic isopods of fishes usually show wide distribution and low host specificity. This study investigated the occurrence of gill parasites in 120 specimens of spotted goatfish Pseudupeneus maculatus, marine fish of great economic importance for fishery community. The fish were captured monthly in the Coast of Pernambuco State, Northeast Brazil, between October 2012 and September 2013. Eleven isopods were found and identified as Rocinela signata. We observed lower infections in the gills of P. maculatus (prevalence 8.3%, mean intensity 1.6±0.3 and mean abundance 0.1±0.3) when compared to other studies of different host fishes from geographically close locations. This parasite has been reported from a wide variety of fish species, not only in Brazil, but also in the eastern Atlantic and the Pacific oceans. This is the first report of R. signata in spotted goatfish.

Chronic granulomatous inflammation in teleost fish Piaractus mesopotamicus: histopathology model study / Inflamación crónica granulomatosa en el pez teleósteo Piaractus mesopotamicus: modelo de estudio histopatológico

ABSTRACT Objective. This study evaluated the cell kinetic and formation of granuloma during chronic inflammation induced by Bacillus Calmette-Guérin (BCG) in the skeletal muscle of Piaractus mesopotamicus, as a histopathology model to study innate immunity. Materials and methods. Sixty fish were divided in two groups: BCG-inoculated and non-inoculated fish and the inflammatory response analyzed 3, 7, 14, 21 and 33 days post-inoculation (DPI) by histopathology after hematoxylin-eosin and Ziehl-Neelsen staining. Results. 3 DPI of BCG showed a diffuse inflammatory reaction mostly composed by mononuclear cells. The inflammation continued diffuse 7 DPI initiating the cellular organization surrounding the inoculum and have continued at 14 DPI with discrete presence of epithelioid-like type cells with acidophilic cytoplasm and floppy chromatin. Higher cellular organization (21 DPI) surrounding the granuloma with intense peripheral mononuclear inflammatory infiltrate and nevertheless, an increase in the number of fibroblasts and macrophage-like cells was observed. The inflammatory process became less diffuse 33 DPI with formation of small amount of granuloma surrounded by the same type of reaction found in bigger granuloma. Both the young and old granuloma presented typical characteristic around the inoculum composed by a layer of epithelioid-like type cells, besides macrophages, some lymphocytes and abundant fibroblasts. Conclusions. This study showed the feasibility in the use of pacus to study chronic granulomatous inflammatory response induced by BCG, characterized by changes in the kinetics of inflammatory cells in skeletal muscle classifying as immune-epithelioid type, similar to granulomatous inflammation caused by M. marinum in teleost fish.

RESUMEN Objetivo. Este estudio evaluó la cinética celular y la formación de granuloma durante la inflamación crónica inducida por el Bacilo Calmette-Guérin (BCG) en el músculo esquelético de Piaractus mesopotamicus, como modelo histopatológico para estudiar la inmunidad innata. Materiales y métodos. Sesenta peces fueron divididos en dos grupos: peces inoculados con BCG y no inoculados y la respuesta inflamatoria analizada en 3, 7, 14, 21 y 33 días post-inóculo (DPI) por medio del análisis histopatológico y tinciones de hematoxilina-eosina y Ziehl-Neelsen. Resultados. 3 DPI de BCG se observó reacción inflamatoria difusa principalmente formada por infiltrado celular mononuclear. Al 7° DPI la inflamación continuaba difusa con inicio de organización celular alrededor del inoculo, que se observó hasta el 14° DPI con discreta presencia de células de tipo epiteliodes con citoplasma acidófilo y cromatina laxa. Para el 21° DPI se observó alta organización celular alrededor del granuloma con intenso infiltrado mononuclear periférico e incremento en el número de fibroblastos y macrófagos. El proceso inflamatorio se tornó menos difuso a los 33 DPI con formación de pequeños granulomas contenidos dentro de uno más grande. Los granulomas formados más rápidamente así como los formados tardíamente, presentaron características típicas alrededor del inóculo compuesta por una camada de células tipo epitelioides, macrófagos, linfocitos y fibroblastos. Conclusiones. Este estudio mostró la viabilidad del uso del P. mesopotamicus para estudiar la respuesta inflamatoria crónica granulomatosa inducida con BCG, caracterizado por la evolución de la cinética de células inflamatorias en el músculo esquelético clasificándolo como de tipo inmune-epitelioide, similar a la inflamación granulomatosa causada por M. marinum en peces teleósteos.

Diversity and Universality of Endosymbiotic Rickettsia in the Fish Parasite Ichthyophthirius multifiliis.

Although the presence of endosymbiotic rickettsial bacteria, specifically Candidatus Megaira, has been reported in diverse habitats and a wide range of eukaryotic hosts, it remains unclear how broadly Ca. Megaira are distributed in a single host species. In this study we seek to address whether Ca. Megaira are present in most, if not all isolates, of the parasitic ciliate Ichthyophthirius multifiliis. Conserved regions of bacterial 16S rRNA genes were either PCR amplified, or assembled from deep sequencing data, from 18 isolates/populations of I. multifiliis sampled worldwide (Brazil, Taiwan, and USA). We found that rickettsial rRNA sequences belonging to three out of four Ca. Megaira subclades could be consistently detected in all I. multifiliis samples. I. multifiliis collected from local fish farms tend to be inhabited by the same subclade of Ca. Megaira, whereas those derived from pet fish are often inhabited by more than one subclade of Ca. Megaira. Distributions of Ca. Megaira in I. multifiliis thus better reflect the travel history, but not the phylogeny, of I. multifiliis. In summary, our results suggest that I. multifiliis may be dependent on this endosymbiotic relationship, and the association between Ca. Megaira and I. multifiliis is more diverse than previously thought.

Rocinela signata (Isopoda: Aegidae) parasitizing the gills of the spotted goatfish Pseudupeneus maculatus (Actinopterygii: Mullidae) in Northeastern Brazil

ABSTRACT Parasitic isopods of fishes usually show wide distribution and low host specificity. This study investigated the occurrence of gill parasites in 120 specimens of spotted goatfish Pseudupeneus maculatus, marine fish of great economic importance for fishery community. The fish were captured monthly in the Coast of Pernambuco State, Northeast Brazil, between October 2012 and September 2013. Eleven isopods were found and identified as Rocinela signata. We observed lower infections in the gills of P. maculatus (prevalence 8.3%, mean intensity 1.6±0.3 and mean abundance 0.1±0.3) when compared to other studies of different host fishes from geographically close locations. This parasite has been reported from a wide variety of fish species, not only in Brazil, but also in the eastern Atlantic and the Pacific oceans. This is the first report of R. signata in spotted goatfish.

A new species of Trichodina Ehrenberg, 1830 (Ciliophora: Trichodinidae) from Rhamdia quelen (Siluriformes: Heptapteridae) and Gymnotus sp. (Teleostei: Gymnotidae) in Brazil.

The present study describes a new species of Trichodina found in the mucus of the body surface of 137 farmed and wild silver catfish (Rhamdia quelen) located in southern Brazil and in the gills of 99 banded knife fish (Gymnotus sp.) captured in the basin of Pantanal. Smears of the body surface mucus and gills were air dried at room temperature and impregnated with silver nitrate and stained with Giemsa for analysis of the adhesive disc, denticles and macronucleus. Characterized as a medium trichodinid, Trichodina quelenii n. sp. resembles T. heterodentata and T. acuta. It differs from T. heterodentata by the shape of the blade and from T. acuta by the lack of central circle. Furthermore, the distinct shape of the blade of T. quelenii n. sp. differs from all previously described species for the genus, leaving no doubt it is a new species.

Hemato-immunological and biochemical parameters of silver catfish Rhamdia quelen immunized with live theronts of Ichthyophthirius multifiliis.

This study investigated the immunization by intraperitoneal injection (i.p.) (assay I) and immersion bath (assay II) with live theronts of Ichthyophthirius multifillis in Rhamdia quelen and its influence on the hemato-immunological and biochemical parameters. Fish were divided in control (non immunized no challenged); non immunized and challenged with 12,000 theronts/fish; non immunized and challenged with 22,000 theronts/fish; immunized and challenged with 12,000 theronts/fish; immunized and challenged with 22,000 theronts/fish. Six days after challenge, either in the assay I or in the assay II the prevalence of I. multifillis in the gills was higher in non immunized fish (33.33% and 27.77%, respectively). In the assay I showed higher numbers of thrombocytes, leukocytes, lymphocytes, neutrophils and monocytes 20 days after injection and lower numbers after challenge. The immunoglobulin values were higher in fish non immunized. Fish immunized by immersion bath (assay II) showed greater values of catalase (CAT) in the liver (1245.49 U/mgprt) when compared to i.p. (198.79 U/mgprt). The levels of CAT in the liver of fish from the assay II were greater (1738.47 U/mgprt) 14 days after immunization than that observed 21 days after (1114.26 U/mgprt). The vaccination by i.p method showed influence on the hematological parameters. On the other hand, the immersion bath vaccination showed greater influence on the catalase activity in the liver. The results showed that new parameters like total protein, immunoglobulin and antioxidant enzymes could be considered in evaluating the host response to infection.

Detection of a parasitic amoeba (Order Dactylopodida) in the female gonads of oysters in Brazil.

The impacts of oocyte parasites on the reproductive success of molluscs are largely unknown. In this study, we evaluated the presence of gonad parasites in 6 species of marine bivalve molluscs native to southern Brazil. Cultured bivalves included the mangrove oyster Crassostrea gasar (sometimes called C. brasiliana), the brown mussel Perna perna, the lion's paw scallop Nodipecten nodosus and the wing pearl oyster Pteria hirundo. Another species of mangrove oyster, C. rhizophorae, and the carib pointed venus clam Anomalocardia brasiliana (syn. A. flexuosa) were collected from the wild. Molluscs were collected in winter 2009 and summer 2010 for histopathological and molecular evaluation. An unknown ovarian parasite (UOP) was observed in histopathological sections of female gonads of C. gasar and C. rhizophorae. The UOP possessed features suggestive of amoebae, including an irregular outer membrane, frothy cytoplasm, a nucleus with a prominent central nucleolus and a closely associated basophilic parasome. PCR analysis was negative for Marteilioides chungmuensis, Perkinsus spp. and Paramoeba perurans. However, real-time PCR successfully amplified DNA from oyster gonads when using universal Paramoeba spp. primers. Also, conventional PCR amplified DNA using primers specific for Perkinsela amoebae-like organisms (syn. Perkinsiella), which are considered as endosymbionts of Parameoba spp., previously thought to be the parasome. Our results suggest that this UOP is a species of amoeba belonging to 1 of the 2 families of the order Dactylopodida, possibly related to Paramoeba spp. This study represents the first report of this type of organism in oysters. We found that C. gasar and C. rhizophorae were the most susceptible molluscs to these UOPs.

Hematological and histopathological changes in silver catfish Rhamdia quelen (Siluriformes) exposed to clomazone herbicide in the Madre River, Santa Catarina State, Southern Brazil.

This study evaluated the influence of the clomazone herbicide (2-[(2-chlorophenyl)methyl]-4,4-dimethyl-3-isoxazolidinone) contamination on the hematological parameters and histological changes in gills and liver of silver catfish (Rhamdia quelen) from Madre River, Santa Catarina State, Southern Brazil. Fish were collected between March 2010 and January 2012 at two different sites of the Madre River, one site receiving residual water (contaminated site) from rice culture (n=49) and another that do not receive residual water (reference site) (n=48). The herbicide clomazone analysis detected 3.40±1.70 µg/L in the contaminated site and 1.1±0.33 µg/L in the reference site. Fish from contaminated site showed increased (P<0.05) number of monocytes suggesting the possible defense response as a result of chronic exposure to clomazone. On the other hand, no difference was found in the hematocrit percentage, red blood cell count, total thrombocyte number, white blood cell count, lymphocytes, and neutrophils number. Fish from both sites showed histopathological changes in gills and liver, possibly caused by chronic exposure to contamination. The influence of herbicide sub doses on fish health is also discussed.

Hematological parameters of Iranian cichlid Iranocichla hormuzensis: Coad, 1982 (Perciformes) in Mehran river.

This study describes the hematological parameters in Iranocichla hormuzensis, an Iranian freshwater cichlid important as ornamental and food fish. Forty fish were captured with seine net at Mehran river Hormozgan province, Iran. Blood was used to determine the total counts of red blood cells (RBC) and white blood cells (WBC), hematocrit, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) and morphometric data of erythrocytes. The Iranian fish showed lower RBC and WBC values than the other cichlids (Oreochromis niloticus, O. aureus, O. mossambicus, O. hybrid, Cichlasoma dimerus and Cichla monoculus). Hematocrit did not vary among the species, but MCV, MCH and MCHC in I. hormuzensis were higher than those for O. niloticus, O. aureus, O. hybrid, C. dimerus and C. monoculus. These differences may be related to different life habit of fish. This study suggests that I. hormuzensis is well acclimated to the environment being the first report for its hematology. It is also suggested high efficiency in oxygen transportation, and an efficient inflow of oxygen by the gills, indicating the welfare of fish on this environment.

Effect of Ichthyophthirius multifiliis parasitism on the survival, hematology and bacterial load in channel catfish previously exposed to Edwardsiella ictaluri.

The effect of Ichthyophthirius multifiliis (Ich) parasitism on survival, hematology and bacterial load in channel catfish, Ictalurus punctatus, previously exposed to Edwardsiella ictaluri was studied. Fish were exposed to E. ictaluri 1 day prior to Ich in the following treatments: (1) infected by E. ictaluri and Ich at 2,500 theronts/fish; (2) infected by E. ictaluri only; (3) infected by Ich at 2,500 theronts/fish only; and (4) non infected control. Mortality was significantly higher in fish previously exposed to E. ictaluri and then infected by Ich (71.1 %). Mortalities were 26.7 %, 28.9 % and 0 % for fish infected by E. ictaluri only, by Ich only and non-infected control, respectively. Quantitative polymerase chain reaction demonstrated the presence of E. ictaluri in the brain, gill, kidney and liver of fish infected with E. ictaluri regardless of Ich parasitism. At day 8, E. ictaluri parasitized fish had significantly more bacteria present in the brain, gill and liver, with no bacteria detected in these organs in the E. ictaluri-only treatment, suggesting that the bacteria persisted longer in parasitized fish. Decreased red blood cells count and hematocrit in fish at days 8 and 19 after co-infection suggests chronic anemia. Lymphocyte numbers significantly decreased in all infected treatments versus the non-infected controls at days 2, 8 and 19. Lymphopenia suggests that lymphocytes were actively involved in the immune response. Bacterial clearance was probably influenced by the stress of parasitism and/or the mucosal response induced by ectoparasitic Ich that resulted in the higher mortality seen in the co-infected treatment.

Enhanced susceptibility of channel catfish to the bacterium Edwardsiella ictaluri after parasitism by Ichthyophthirius multifiliis.

Bacterium Edwardsiella ictaluri and parasite Ichthyophthirius multifiliis (Ich) are two common pathogens of cultured fish. The objective of this study was to evaluate the susceptibility of channel catfish Ictalurus punctatus to E. ictaluri and determine bacterial loads in different fish organs after parasitism by Ich. Fish received the following treatments: (1) infected by I. multifiliis at 5000 theronts/fish and exposed to E. ictaluri; (2) infected by I. multifiliis alone; (3) exposed to E. ictaluri alone; and (4) non-infected control. E. ictaluri in fish organs were quantified by quantitative real-time polymerase chain reaction and reported as genome equivalents per mg of tissue (GEs/mg). The results demonstrated that the Ich-parasitized catfish showed significantly (P<0.05) higher mortality (91.7%) when exposed to E. ictaluri than non-parasitized fish (10%). The bacterial loads in fish infected by 5000 theronts/fish ranged from 6497 to 163,898 GEs/mg which was between 40 and 2000 fold higher than non-parasitized fish (49-141 GEs/mg). Ich infection enhanced the susceptibility of channel catfish to bacterial invasion and increased fish mortality.