Extranodal gammadelta-T-cell lymphoma in a dog with leishmaniasis

An 8-year-old intact male mongrel dog with alopecia and weight loss was referred to the Veterinary Faculty of Naples. The dog had pale mucous membranes, enlarged prescapular lymph nodes, and splenomegaly. Laboratory abnormalities included anemia, thrombocytopenia, and hyperglobulinemia. Bone marrow aspirate smears contained numerous Leishmania amastigotes and an immunofluorescent antibody titer was strongly positive (1:1280) for leishmaniasis. The dog was treated with a combination of meglumine antimoniate and allopurinol for 60 days and showed clinical improvement. Two months after the end of treatment the dog was again referred because of relapse of leishmaniasis and the presence of a firm subcutaneous mass on the medial right thigh. Based on cytologic examination of fine needle aspirates of the mass, a diagnosis of large-cell lymphoma was made. Flow cytometry of tumor cells revealed gammadelta-T-cell lymphoma with a CD5+, CD3+, TCRgammadelta+, CD4-, CD8-, CD45RA+ immunophenotype. Using nested PCR, amastigotes were not detected in the neoplastic tissue. An association between leishmaniasis and hematopoietic tumors has been described rarely. gammadelta-T cells may be involved in the host response to this parasite, and prolonged antigenic stimulation and chronic immunosuppression (typical of leishmaniasis) play a crucial role in the etiopathogenesis of T-cell lymphoma.     

Isolation and detection of Fasciola hepatica DNA in Lymnaea viatrix from formalin-fixed and paraffin-embedded tissues through multiplex-PCR

Detection of Fasciola hepatica infection in Lymnaea viatrix through analysis of histological cuts is based upon morphological characters of the parasite during the intra-mollusk phase of parasitism. At this stage, trematode forms are very similar and, thus, very difficult to differentiate. Specific detection may also be impaired by the presence of other helminthes in the mollusk. Histological samples are usually fixed in formalin, embedded in paraffin, sectioned and HE stained. In the current study, a method for the extraction of DNA from formalin-fixed, paraffin-embedded tissues was standardized by means of deparaffinizing with xylol and digesting with proteinase K. Extracted DNA was amplified in a multiplex-PCR, by using simultaneous primers in a single reaction under high stringency conditions. Results showed specific amplification of DNA from the trematode and the snails. The technique was sensitive enough to detect F. hepatica infections in L. viatrix, in histological sections in which the presence of larval stages could not be observed through brightfield microscopy. The profiles generated were: stair bands referring to F. hepatica DNAmt amplification; a band of 1200 bp referring to L. viatrix ITS and another of 1300 bp referring to F. hepatica ITS and other trematodes. Multiplex-PCR has shown to be a fast, safe, highly sensitive and specific method, which is able to amplify DNA from fixed tissues, despite a low DNA quantity and its degradation caused by fixation processes. Such methodology may be useful in studies on fascioliasis epidemiology, enabling the use of material from histological collections.     

Clinical value of anti-Leishmania (Leishmania) chagasi IgG titers detected by flow cytometry to distinguish infected from vaccinated dogs

Leishmune vaccination covers a broader number of endemic areas of canine visceral leishmaniasis (CVL) and therefore the development of new serological devices able to discriminate CVL from Leishmune vaccinees becomes an urgent need considering the post-vaccine seroconversion detected throughout conventional methodologies. Herein, we have described the establishment of a flow cytometry based methodology to detect anti-fixed L. (L.) chagasi promastigotes antibodies (FC-AFPA-IgG, FC-AFPA-IgG1 and FC-AFPA-IgG2) in sera samples from Leishmania (Leishmania) chagasi infected dogs and Leishmune vaccinees. The results of FC-AFPA were reported along the sera titration curve (1:128-1:524,288), as percentage-of-positive-fluorescent-parasite (PPFP). The use of PPFP=20% as a cut-off edge to segregate negative and positive results at sera dilution 1:2048 revealed outstanding performance indexes that elect FC-AFPA-IgG and IgG2 (both detected by polyclonal FITC-labeled second step reagent) applicable to the serological diagnosis of CVL, with 100% of specificity for both IgG and IgG2 and 97 and 93% of sensitivity, respectively. Moreover, FC-AFPA-IgG, applied at sera dilution 1:2048, also appeared as a useful tool to discriminate L. chagasi infected dogs from Leishmune vaccinees, with 76% of specificity. Outstanding likelihood indexes further support the performance of FC-AFPA-IgG for exclusion diagnosis of CVL in Leishmune vaccinees. Analysis of FC-AFPA-IgG at sera dilution 1:8192 revealed the most outstanding indexes, demonstrating that besides the ability of PPFP 相似文献   

Neurotoxic Shellfish Poisoning

Neurotoxic shellfish poisoning (NSP) is caused by consumption of molluscan shellfish contaminated with brevetoxins primarily produced by the dinoflagellate, Karenia brevis. Blooms of K. brevis, called Florida red tide, occur frequently along the Gulf of Mexico. Many shellfish beds in the US (and other nations) are routinely monitored for presence of K. brevis and other brevetoxin-producing organisms. As a result, few NSP cases are reported annually from the US. However, infrequent larger outbreaks do occur. Cases are usually associated with recreationally-harvested shellfish collected during or post red tide blooms. Brevetoxins are neurotoxins which activate voltage-sensitive sodium channels causing sodium influx and nerve membrane depolarization. No fatalities have been reported, but hospitalizations occur. NSP involves a cluster of gastrointestinal and neurological symptoms: nausea and vomiting, paresthesias of the mouth, lips and tongue as well as distal paresthesias, ataxia, slurred speech and dizziness. Neurological symptoms can progress to partial paralysis; respiratory distress has been recorded. Recent research has implicated new species of harmful algal bloom organisms which produce brevetoxins, identified additional marine species which accumulate brevetoxins, and has provided additional information on the toxicity and analysis of brevetoxins. A review of the known epidemiology and recommendations for improved NSP prevention are presented.     

Water quality,Vibrio density and growth of Pacific white shrimp Litopenaeus vannamei (Boone) in an integrated biofloc system with red seaweed Gracilaria birdiae (Greville)

An indoor trial was conducted for 42 days to evaluate water quality, Vibrio density and growth of Litopenaeus vannamei in an integrated biofloc system (IBS) with Gracilaria birdiae. Four treatments were used, each in triplicate: Control (monoculture shrimp); IBS 2.5 (L. vannamei and 2.5 kg wet weight seaweed m^?3); IBS 5.0 (L. vannamei and 5.0 kg wet weight seaweed m^?3) and IBS 7.5 (L. vannamei and 7.5 wet weight seaweed m^?3). Shrimp individuals (0.34 ± 0.01 g) were stocked at a density of 500 shrimp m^?3. No water exchange was carried out during the experimental period. Molasses was added once a day as an organic carbon source to maintain the C:N ratio at 12:1. The IBS significantly decreased (P < 0.05) dissolved inorganic nitrogen (DIN) ranging from 19% to 34% (3.12–3.83 mg L^?1), NO₃‐N ranging from 19% to 38% (2.40–3.16 mg L^?1), Vibrio density ranging from by 8–83% (0.40–2.20 log 10³ colony‐forming units mL^?1), and FCR ranging from by 20–30% (1.20–1.37), as compared to the control (4.73 mg L^?1, 3.93 mg L^?1, 2.40 log 10³ colony‐forming units mL^?1, and 1.74 respectively). Moreover, the IBS significantly increased (P < 0.05) crude protein in whole body shrimp, ranging from 8% to 13% (13.2–13.7% wet weight basis); as well as final weight, ranging from 25% to 32% (3.90–4.12 g), weekly growth ranging from 25% to 34% (0.59–0.63 g), and shrimp yield by 22–39% (1.72–1.96 kg m^?3), as compared to control (12.1% wet weight basis, 3.12 g, 0.47 g, and 1.41 kg m^?3 respectively). It can thus be concluded that cultivating Gracilaria birdiae in an IBS with L. vannamei can contribute to DIN and NO₃‐N removal, lower Vibrio density, increased crude protein in whole body shrimp, higher growth and yield parameters in shrimp culture.     

Intensive fishing has not forced dietary change in the South American fur seal Arctophoca (=Arctocephalus) australis off Río de la Plata and adjoining areas

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Effect of feeding frequency on growth and digestive enzyme activity in Litopenaeus vannameiduring the grow‐out phase in biofloc system

The present study evaluated the effects of feeding frequency during the grow‐out phase of Litopenaeus vannameiin biofloc system. The experiment comprised of four treatments with three replicates each, corresponding to the following feeding frequencies: one (8:00 a.m.), two (8:00 a.m. and 4:00 p.m.), three (8:00 a.m., 12:00 p.m. and 4:00 p.m.) and four times a day (8:00 a.m., 10:00 a.m., 12:00 p.m. and 4:00 p.m.). Zootechnical performance and digestive enzyme activity were monitored for 63 days. At the end of the experiment, the highest survival rates were found for the individuals fed three and four times a day. Wet weight gain (6.63 g) and specific growth rate (3.46% per day) of the individuals fed three times a day were significantly higher compared to animals fed one and four times a day, but did not differ significantly to the group fed twice a day (6.15 g and 3.46% per day, respectively). The digestive enzyme activities were not affected by the feeding frequency, but negative correlations were found between these activities and shrimp weight. It is recommended that L. vannamei should be fed three times a day during the grow‐out phase in a biofloc system under the conditions used in this study.     

Olive oil by-product as a natural antioxidant in gilthead sea bream (<Emphasis Type="Italic">Sparus aurata</Emphasis>) nutrition

The aim of the study reported here was to evaluate the efficiency of a natural antioxidant substance in gilthead sea bream (Sparus aurata) feeds. An olive oil by-product, olive mill vegetation water (VW), contains polyphenols, which have a strong antioxidant activity. A 147-day growth trial was conducted (monofactorial balanced, 4 × 3) with diet as the experimental factor. Two diets [isonitrogenous (crude protein 40%) and isoenergetic (gross energy = 18MJ kg⁻¹ on a dry weight basis] were formulated with 1 and 5% of VW (VW1, VW5); the control diet did not contain VW. A total of 600 juvenile gilthead sea breams (mean body weight 114.1 ± 5.7 g) were utilised. Production parameters and somatic indexes were calculated at the end of the growth trial. Antioxidant activity in fish fillets was investigated using TBARS and DPPH assays. A number of haematological parameters and digestive enzyme patterns were measured in fish in the middle and at the end of the experiment. The TBARS values showed slight delays in the development of oxidation in the fillet of fish fed with VW. There was no statistical difference between fish fed with the experimental diet and the control group, except for maltase activity, which increased with increasing VW in the feed. We found that the use of VW in a gilthead sea bream diet did not have any detrimental effects on gilthead sea bream production and physiological parameters and slightly improved the conservation of the fish fillets.