Comparison of the susceptibility of brown trout (Salmo trutta) and four rainbow trout (Oncorhynchus mykiss) strains to the myxozoan Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD)

Differences in susceptibility to the myxozoan parasite Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD), between four strains of rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta) were evaluated. Fish were exposed to water enzootic for the parasite in the field for 5 days and were subsequently transferred to the laboratory. Relative parasite load was determined after 2, 3 and 4 weeks post-exposure (wpe) by quantitative real-time PCR (qPCR) of kidney samples and number of parasite stages was determined in immunohistochemical stained sections of kidney, liver and spleen tissues. According to qPCR results, the highest amount of parasite DNA per equal amount of host tissue at all time points was measured in brown trout. Two of the rainbow trout strains showed lower relative parasite load than all other groups at the beginning of the experiment, but the parasite multiplied faster in these strains resulting in an equal level of relative parasite load for all rainbow trout strains at 4 wpe. A weak negative correlation of fish size and parasite load was detected. Only in samples of a few fish, single stages of T. bryosalmonae were found in sections stained by immunohistochemistry impeding quantitative evaluation of parasite numbers by this method. The results indicate a differential resistance to T. bryosalmonae between the rainbow trout strains investigated and between rainbow trout and brown trout.     

Molecular regulation of lipid metabolism in liver and muscle of rainbow trout subjected to acute and chronic insulin treatments

Although the metabolic actions of insulin in fish have been investigated widely in the past several years, lipid metabolism has received little attention, especially in tissues like the liver or white muscle. In the present study, rainbow trout received insulin treatments both acutely (intraperitoneal injection) and chronically (through mino-osmotic pumps) to elucidate hormone metabolic actions at molecular levels on the 2 main insulin target tissues in trout, namely, liver and muscle. Plasma and free fatty acid concentrations in plasma, as well as mRNA measurements of some key enzymes involved in lipid metabolism, were assessed in these tissues after 6 h and 4 d of acute and chronic insulin treatments, respectively. Our results showed that although fish received the same final total amount of hormone in both treatments, the actions of insulin on lipid metabolism were both time and tissue dependent. After the acute insulin treatment, the main anabolic role of insulin was reflected in decreased plasma free fatty acid concentrations linked to enhanced hepatic lipogenesis. We also found that insulin increased the mRNA levels of enzymes involved in lipid oxidation, perhaps to counteract insulin-induced hypoglycemia. In contrast, our data show that after chronic insulin treatment, liver and muscle exhibit different metabolic strategies: whereas in the liver chronic insulin-induced hypoglycemia may stimulate lipolytic processes to spare glucose stores, the muscle responds directly to the anabolic hormone action by increasing its lipogenic capacity and by inhibiting pathways of lipid oxidation.     

Myxobolus cerebralis infection in rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta) exposed under natural stream conditions.

From early April into mid-June 1977, sequential groups of juvenile rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta) were each exposed for 10 days to the parasite Myxobolus cerebralis by immersion in a stream inhabited by infected wild trout. Following incubation in a M. cerebralis-free facility, trout were subsequently killed, and heads and gill arches were examined by routine histologic methods. A grading scale to quantify lesion severity was developed and applied. Percentage infected, lesion severity scores, effects of water temperature and flow rates on percentage infected and lesion severity scores, and resulting pathology were determined for each species at each exposure period. The percentage of rainbow trout infected with M. cerebralis was significantly higher than the percentage of brown trout infected for each exposure period. The percentages of rainbow trout infected in exposure periods later in the calendar year were significantly higher than those in earlier periods. The percentages of brown trout infected were not significantly different among exposure periods. Overall average lesion severity scores were significantly higher in rainbow than in brown trout. Lesion severity scores in rainbow trout increased over time (a positive correlation with exposure period). Lesion severity scores were not significantly different for brown trout among exposure periods. A significant correlation existed between water temperature and percentage of rainbow trout infected; a significant correlation also existed between water temperature and lesion severity scores in rainbow trout. Similar correlations did not exist for percentage of brown trout infected or accompanying lesion severity scores. In rainbow trout, ventral calvarium was the most common site of M. cerebralis replication, followed by gill arches. In brown trout, lesions were virtually confined to gill arches. Early lesions consisted of foci of cartilage necrosis with small numbers of M. cerebralis developmental stages. More advanced lesions consisted of multifocal areas of cartilage necrosis with numerous M. cerebralis developmental stages and/or mature myxospores bordered and/or infiltrated by mono- and multinuclear leukocytes. Lesions in brown trout were smaller and had fewer associated leukocytes and M. cerebralis developmental stages and/or mature myxospores. Higher infection rates, lesion severity scores, and differences in lesion location in rainbow versus brown trout explain in part why numbers of rainbow but not brown trout have fallen in western rivers inhabited with M. cerebralis-infected trout.     

Prevalence of vancomycin-resistant Enterococcus faecium (VREF) in pig faeces from slaughterhouses in Spain

A study to estimate the prevalence of vancomycin-resistant Enterococcus faecium in faecal samples from pigs at slaughterhouses in Spain was carried out between November 1998 and January 1999 with 900 samples taken from four abattoirs representing 9.7% of all pig slaughtered in 1998. Using a selective enrichment broth with vancomycin (8 μg/ml), 64 samples (7.1%; 95% CI: 5.5, 9.0%) had E. faecium vancomycin-resistant strains that showed minimal inhibitory concentrations of 256 μg/ml (62 strains) and 512 μg/ml (two strains). Results by farm showed that 43 of the 240 pig farms represented in the sampling had at least one faecal sample with vancomycin-resistant E. faecium.     

The binding of soybean agglutinin (SBA) to the intestinal epithelium of Atlantic salmon, Salmo salar and rainbow trout, Oncorhynchus mykiss, fed high levels of soybean meal.

A lectin present in soya, soybean agglutinin (SBA), was identified in electrophoretic profiles and immunoblots of dehulled solvent-extracted soybean meal (DSSM), full-fat soybean meal (FFSM) and of aqueous extracts of feeds incorporating them in their formulation. A quantitative estimation was made of the proportion of SBA comprising the total protein in FFSM and a trial diet was prepared containing an amount of pure SBA similar to that in diets incorporating high levels of the whole soya product. Fish fed with this diet exhibited similar pathological disruption of the intestinal tract to that observed in fish given a diet with a high level of DSSM (60% of the diet). Furthermore, immuno-histochemistry revealed the binding of the SBA to the enterocytes lining the intestinal villi both of fish fed a diet incorporating pure SBA and those fed a diet containing a high-level of soya (60%). Our results suggest that SBA binds in vivo to the intestinal epithelium of fish and has a contributory role in pathological changes associated with fish feeds containing high levels of soybean proteins.     

Intestinal and haematic parasitism in the birds of the Almuñecar (Granada, Spain) ornithological garden

Birds from the Almuñecar ornithological garden (Granada, Spain) were surveyed from June 2006 to May 2007 to establish programmes to prevent, control, and treat intestinal and haematic parasites. A total of 984 faecal samples and 41 samples of blood were collected from Psittacidae, Cacatuidae, Phasianidae, and Anatidae. One or more intestinal parasites were identified in 51.6% of the samples. Blood parasites were found in 26.8% of the birds examined. The most frequent pathogenic endoparasites were coccidians, such as Cyclospora sp. (4.5%), Eimeria sp. (4.1%) and Isospora sp. (2%) and helminths such as Capillaria sp. (10. 1%), Ascaridia sp. (4.9%) and Heterakis gallinarum (4.9%). All the parasites varied with season but the most were found year round. Multiple parasitic infections by intestinal parasites were common, with 196 of 984 faecal samples having 2–5 intestinal parasites. The most frequent cases of multiple parasitism were Blastocystis plus Entamoeba sp. and Blastocystis plus Cyclospora sp. The haematic protozoa detected were Haemoproteus sp. (17%) and Plasmodium sp. (7.3%). Multiple parasitism by Haemoproteus sp. and Plasmodium sp. was detected in 1 sample of Gallus gallus. After each sampling, some of the affected animals were treated according to our results, and the corresponding programmes of prevention and control were designed.     

Residues of clenbuterol in tissues of the rainbow trout (Oncorhynchus mykiss)

There have been many studies on the efficacy of ₂-adrenergic drugs as feed additives but no data are available at present on the use of clenbuterol in fish production. To evaluate the residues of clenbuterol in tissues of fish, 50 trout (Oncorhynchus mykiss) were fed for 21 days on a fish feed containing 5 ppm of the drug. The livers, muscles and skins of sample groups of fish were analysed by HPLC with visible spectrophotometric detection on days 15 and 21 of treatment and at intervals during a 30-day withdrawal time. Clenbuterol reached its highest levels in the liver (mean 440 ppb; SD=±159;n=5) on day 15 of treatment, with a slow depletion curve; 24±3 ppb was still present at the end of the withdrawal period. At this time, residues were still present in the edible tissues, i.e. muscle (5±1 ppb) and skin (7±3 ppb). Side-effects were noted during the first week of treatment.Abbreviations ppb parts per billion - BHT butylated hydroxy toluene - CV coefficient of variation - DAD diode array detector - HPLC high-performance liquid chromatography     

An outbreak of white spot disease (Ichthyophthirius multifiliis) in young fingerling rainbow trout (Salmo gairdneri Richardson)

An outbreak of white spot disease (‘Ich’) in fingerling rainbow trout is reported. Approximately 3,000 fish died or were killed in extremis in a period of 5 weeks. Post mortem examinations were carried out on 12 fish, all of which showed thickened whitish foci on the skin. One fish showed skin ulceration and another gill ulcerations. The gill infestation was more severe than the skin. Pathological results are described and discussed.     

Host and environmental risk factors associated with Cryptosporidium scophthalmi (Apicomplexa) infection in cultured turbot, Psetta maxima (L.) (Pisces, Teleostei)

An epidemiological cohort study of Cryptosporidium scophthalmi in cultured turbot Psetta maxima L. of Northwestern Spain was conducted along a four-year period. Four different ongrowing cohorts were monitored monthly from introduction into the ongrowing tanks (10–50 g) until reaching market size (400–1400 g). The association of host and environmental factors with five categories of parasite abundance was assessed using a multivariable regression framework. Epidemiological factors assessed here were water temperature, weight, length, month of collection, season, age, origin, condition factor, water filtration, and status to the myxozoan Enteromyxum scophthalmi infection. E. scophthalmi was included into the analysis because it targets the same organ than C. scophthalmi and it was prevalent in the studied population. The multivariable analysis demonstrated the statistically significant association between several factors and parasite abundance. C. scophthalmi abundance was associated (P < 0.05) with age, condition factor, season, and status to E. scophthalmi infection. Young animals, with poor condition factor, during spring or summer, and not infected with the myxozoan were most likely to be highly infected by C. scophthalmi. Inclusion of these four variables significantly (P < 0.05) improved the model, compared to the model that did not include any of these epidemiological factors. Increasing levels of C. scophthalmi abundance were associated (P < 0.01) with higher severity of C. scophthalmi-compatible lesions. The frequency of distribution of C. scophthalmi abundance was clearly right-skewed and fitted a negative binomial distribution, whereas the intensity of infection fitted a Poisson distribution. The quantification of the variance-to-mean ratio stratified by age demonstrated overdispersion for 8–16 months old fish, although this bivariate association is likely affected by several other factors, as suggested by the results of the multivariable analysis. The negative relation between C. scophthalmi abundance and status to E. scophthalmi infection suggests differences in the transmission, onset, and course of both infections. The coarse filtration used in some cohorts did not significantly affect the levels of infection. C. scophthalmi was probably introduced into the ongrowing tanks mainly with carrier fish, though the involvement of infective oocysts from the water supply cannot be disregarded. Infection prevalence and mean intensity decreased with fish age and a seasonal distribution was found. Results presented here will help to understand the epidemiology of C. scophthalmi in turbot, to estimate the expected levels of infection associated with presence or absence of epidemiological factors, and to quantify the impact that the disease may have on susceptible turbot populations. The multivariable model used here is more powerful than the visual inspection of graphics for exploring associations in cooperative processes and can be easily extended to the assessment of epidemiological associations in other population and parasitic diseases.     

Genetic typing and prevalence of Border disease virus (BDV) in small ruminant flocks in Spain

Between 2001 and 2002, samples from 1,413 animals in 21 Spanish small ruminant flocks, most of them with animals showing clinical signs compatible with Border disease (BD), were screened for the presence of Pestivirus antigen and antibodies by an indirect peroxidase monolayer assay (IPMA) and the virus neutralization test (VNT), respectively. Although all flocks harboured seropositive animals, virus could only be isolated from animals in five of the flocks. Between 4 and 11 months later all animals older than 6 months in three of the flocks were resampled. At this time, 51-83% of them had neutralizing antibodies. The prevalence of persistently infected (PI) animals within two of the flocks was 0.3 and 0.6%, respectively. The third flock presumably had eliminated all the PI animals. Fourteen virus isolates were obtained. The 5' untranslated region (5'UTR) was amplified by RT-PCR and directly sequenced. Phylogenetic analyses classified them as a group of Border disease viruses (BDV), separated from BDV-1, but showing a relatively low bootstrap value. Three of the 14 isolates were in the same subgroup as a set of formerly characterised Spanish isolates from the Basque Country, which were allocated to subgroup BDV-C. In addition, they were in the group with an isolate from chamois, which is currently allocated in group BDV-4. Because of its close relation to the chamois isolate, these isolates were tentatively reallocated in a subgroup BDV-4a. The remaining isolates generated a new subgroup, related but not in the same cluster as the chamois isolate, and was therefore tentatively assigned to a new subgroup BDV-4b. Our results show that classification and nomenclature of BDV needs to be harmonised.     

The prevalence of green muscle disease in turkeys from the Warmia and Mazury province of Poland

The green muscle disease (GMD), described for the first time in 1968, is a cause of significant losses in the production of turkey meat. Pathological changes of different intensity are found in smaller pectoral muscles on one or both sides of the sternal crest. GMD has been more frequently observed in Poland since 1991 in breeding turkeys of heavy and medium-heavy types assigned to slaughter after completing the reproductive period. The present study deals with the occurrence of this disease in the Warmia and Mazury District of Poland. Characteristic green foci located in small pectoral muscles were found in 4090 out of 26,169 slaughtered turkeys, i.e. in 15.63% of the carcasses investigated. It was established that 2430.5 kg of meat were confiscated what gave 0.59 kg of confiscated meat per head on the average.     

在豆粕型饲料中添加蛋氨酸羟基类似物(MHA)对虹鳟鱼生产性能的影响

试验2本试验的目的是研究MHA在豆粕型实用配方中的最适添加量。     

Epidemiology of Oestrus ovis (Linneo, 1761) infestation in goats in Spain

This survey was conducted to determine the chronobiology and seroprevalence of nasal bot infestation (Oestrus ovis) in Spain and to identify the risk factors associated with this disease in caprine herds.

A total of 1590 sera from adult goats were collected at random on 175 farms in southwestern Spain. Sera were tested by ELISA, using crude protein from second stage larvae as antigen. The mean seroprevalence was 46.04% and mean percentage of optical densities was 41.83. These data indicate a high prevalence of this parasite in the investigated areas. The serological survey revealed that goats managed at higher altitudes, at meridians latitudes and on farms with small herds had a smaller probability of infestation. Eighty goat heads, obtained from abattoirs in the central region of Spain, were collected and examined for nasal botflies from February to October 2002. O. ovis larval stages were recovered from the nasal-sinus cavities of 23 goats, reaching a prevalence of 34.94%. The mean larval burden was 3.9 larvae per infested head. No first instars were found during February and March, when the second instar reached its larger count. The third instar was observed in very small number during the whole period of study, with one peak occurring in July–August. These data show the existence of a favourable period for the development of larval instars of O. ovis in goats that starts in February and finishes in September.     

Influenza A Virus Surveillance in the Invasive American Mink (Neovison vison) from Freshwater Ecosystems,Northern Spain

Influenza A viruses (IAVs) are negative‐sense, single‐stranded and segmented RNA viruses of the Orthomyxoviridae family that may cause acute respiratory disease in a wide range of birds and mammals. Susceptibility of several species within the family Mustelidae to IAVs has been reported as a result of natural or experimental infections. The objectives of this study were to assess whether free‐ranging American mink populations from Northern Spain were infected with IAV and try to define the role of this species in the epidemiology of IAV. Sera from 689 American mink from Northern Spain captured between 2011 and 2014 were tested for the presence of antibodies against IAVs using a commercial competition cELISA. Positive sera were further analysed with haemagglutination inhibition (HI) assay. Fifteen of the 689 (2.2%, 1.3–3.6 CI_95%) of the American minks analysed were ELISA positive. No significant differences were observed between years of capture, provinces, river basins, sexes or ages of the animals. All seropositive sera resulted negative to the panel strains used in the HI assay, showing that the most relevant strains circulating in swine, the most relevant avian subtypes (H5 and H7) and the H10N4 subtype isolated in minks have not been circulating in this free‐ranging exotic carnivore from Spain. In the light of these results, the free‐range American mink from Northern Spain do not seem to have an important role in the epidemiology of IAVs.