Evaluation of the growth performance and the welfare status in sea bass (Dicentrarchus labrax) reared under organic and traditional aquaculture by immunohistochemical and biomolecular approach

A growing consumer awareness of food quality, based on high degree of safety, adequate nutritional value, sustainable production, eco-environmental attention, animal welfare and use of raw materials, has facilitated the spread of the "organic production". Organic aquaculture has lagged behind the organic agriculture sector in terms of quantities and diversity of certified organic products because of the absence of detailed accepted standards and criteria until recently. The main challenges for organic aquaculture are to improve the coordination between production and market and to achieve an appropriate framework to drive further development. In organically cultured fish, differences in feeds and nutrition compared to conventional systems are likely to result in differences in the quality of the flesh, and this is a significant factor in consumer choice. The present study aims at evaluating the growing performances, IGF-I and IGF-II levels, oxidative stress and contaminant markers in European sea bass (Dicentrarchus labrax) fed with organic and conventional diet in field conditions. Growing performances and condition factors showed a positive growing trend in both groups, as evidenced by molecular analyses. IGF-I and IGF-II showed similar levels throughout the whole experimental period confirming their role during growth. A greater productivity in conventional fed fish compared to the organic ones was observed. The higher productivity was likely due to diet composition, since differences were significantly mitigated during starvation period. Fillet analysis of organic sea bass showed a higher content in MUFA and lower in PUFAs n-6 indicating that diets with a content in fatty acids closer to that of wild fish reflect the same fatty acid profile of the flesh. On the other hand, the considered oxidative stress and contaminant markers did not show any significant differences among groups

Chromogenic in situ hybridization for the detection of lambda and kappa immunoglobulin light chains as a potential auxiliary diagnostic technique in canine plasmacytomas

The heterogeneous morphologic features of canine plasmacytomas (PCTs) can make their differentiation from other round cell tumors challenging. Immunohistochemistry (IHC) for lambda (\u3bb) and kappa (\u43a) immunoglobulin (Ig) light chains is often equivocal because of high background staining. The chromogenic in situ hybridization (CISH) technique for light chains has shown higher sensitivity compared to IHC in human plasma cell tumors. Therefore, we aimed to validate automated CISH for light chains in canine tissues and to evaluate its diagnostic potential in canine PCTs, in conjunction with routinely used IHC markers. CISH for light chains demonstrated a clear signal in plasma cell populations of canine control tissues (lymph nodes, lymphoplasmacytic inflammation) showing a polyclonal pattern with a prevalence of \u3bb-producing cells. CISH detected monotypic light chain expression in 33 of 53 (62%) PCTs, 31 expressing \u3bb and 2 expressing \u43a. CISH was more sensitive than IHC for \u3bb light chain (58% vs. 47%, respectively) and more easily interpretable given the absence of confounding background staining. The absence of CISH staining for both \u3bb and \u43a in a considerable subset of tumors may be the result of lower light chain production by neoplastic cells. Multiple myeloma oncogene 1 (MUM1) was expressed by all but 2 PCTs (96%), which showed \u3bb expression by CISH and IHC. The identification of poorly differentiated canine PCTs requires the assessment of a panel of IHC markers, with the potential support of CISH for Ig light chains

Frequency of DEA 1 antigen in 1037 mongrel and PUREBREED dogs in Italy

Background: The prevalence of dog erythrocyte antigen (DEA 1) in canine population is approximately 40\u201360%. Often data are limited to a small number of breeds and/or dogs. The aims of this study were to evaluate frequency of DEA 1 in a large population of purebred and mongrel dogs including Italian native breeds and to recognize a possible association between DEA 1 and breed, sex, and genetic and phenotypical/functional classifications of breeds. Frequencies of DEA 1 blood group collected from screened/enrolled blood donors and from healthy and sick dogs were retrospectively evaluated. The breed and the sex were recorded when available. DEA 1 blood typing was assessed by immunocromatographic test on K3EDTA blood samples. The prevalence of DEA 1 antigen was statistically related to breed, gender, F\ue9d\ue9ration Cynologique Internationale (FCI) and genotypic grouping. Results: Sixty-two per cent dogs resulted DEA 1+ and 38% DEA 1-. DEA 1- was statistically associated with Dogo Argentino, Dobermann, German Shepherd, Boxer, Corso dogs, the molossian dogs, the FCI group 1, 2 and 3 and the genetic groups \u201cworking dogs\u201d and \u201cmastiff\u201d. DEA 1+ was statistically associated with Rottweiler, Briquet Griffon Vend\ue9en, Bernese mountain dog, Golden Retriever, the hunting breeds, the FCI group 4, 6, 7 and 8 and the genetic groups \u201cscent hounds\u201d and \u201cretrievers\u201d. No gender association was observed. Conclusions: Data obtained by this work may be clinically useful to drive blood donor enrollment and selection among different breeds

Evaluation of the growth performance and the welfare status in sea bass (Dicentrarchus labrax) reared under organic and traditional aquaculture by immunohistochemical and biomolecular approach

A growing consumer awareness of food quality, based on high degree of safety, adequate nutritional value, sustainable production, eco-environmental attention, animal welfare and use of raw materials, has facilitated the spread of the "organic production". Organic aquaculture has lagged behind the organic agriculture sector in terms of quantities and diversity of certified organic products because of the absence of detailed accepted standards and criteria until recently. The main challenges for organic aquaculture are to improve the coordination between production and market and to achieve an appropriate framework to drive further development. In organically cultured fish, differences in feeds and nutrition compared to conventional systems are likely to result in differences in the quality of the flesh, and this is a significant factor in consumer choice. The present study aims at evaluating the growing performances, IGF-I and IGF-II levels, oxidative stress and contaminant markers in European sea bass (Dicentrarchus labrax) fed with organic and conventional diet in field conditions. Growing performances and condition factors showed a positive growing trend in both groups, as evidenced by molecular analyses. IGF-I and IGF-II showed similar levels throughout the whole experimental period confirming their role during growth. A greater productivity in conventional fed fish compared to the organic ones was observed. The higher productivity was likely due to diet composition, since differences were significantly mitigated during starvation period. Fillet analysis of organic sea bass showed a higher content in MUFA and lower in PUFAs n-6 indicating that diets with a content in fatty acids closer to that of wild fish reflect the same fatty acid profile of the flesh. On the other hand, the considered oxidative stress and contaminant markers did not show any significant differences among groups.La crescente domanda di prodotti alimentari ottenuti da agricoltura biologica in questi ultimi anni è verosimilmente l’espressione di una aumentata consapevolezza del consumatore che sempre più spesso ricerca alimenti di qualità. La qualità del prodotto alimentare derivante da produzioni biologiche non si basa soltanto sul concetto tradizionale di salubrità e di valore nutrizionale del prodotto stesso, ma anche sul concetto globale di sostenibilità e salvaguardia ambientale. L'agricoltura biologica infatti è un metodo di produzione che rispetta il benessere umano e animale e difende la biodiversità ambientale e culturale dei territori proponendo un modello di sviluppo sostenibile volto a salvaguardare ambiente e territorio e valorizzare la qualità delle risorse delle comunità locali. Questo studio di campo si prefigge di confrontare le performances di crescita e il benessere in branzini (Dicentrarchus labrax) alimentati con dieta convenzionale rispetto a branzini alimentati con dieta certificata biologica attraverso un approccio immunoistochimico e biomolecolare valutando i fattori di crescita (IGF-I e IGF-II), parametri dello stress ossidativo e di contaminazione ambientale. Le performances di crescita e i parametri biometrici hanno dimostrato un trend di crescita positivo in entrambi i sistemi di allevamento, come confermato anche dalla quantificazione dei fattori di crescita. I fattori di crescita hanno evidenziato livelli similari in entrambi i gruppi e durante tutto il periodo sperimentale confermando il loro ruolo determinante nella crescita corporea. Il gruppo alimentato con la dieta convenzionale ha mostrato una produttività più elevata rispetto al gruppo biologico. La più elevata produttività registrata a carico del gruppo convenzionale è verosimilmente determinata dalle differenze nella composizione della dieta, considerato che tali differenze venivano mitigate durante i periodi di digiuno. L’analisi del filetto ha evidenziato un più alto contenuto in acidi grassi monoinsaturi e un più basso contenuto in acidi grassi polinsaturi (n- 6) a carico dei filetti biologici Questo dato sta ad indicare che la dieta biologica caratterizzata da un contenuto in acidi grassi più vicino a quello del pesce pescato probabilmente influisce sul profilo degli acidi grassi delle carni di pesce. Per contro, i markers di stress ossidativo e contaminazione ambientali considerati in questo studio di campo non hanno dimostrato alcuna differenza significativa tra i due gruppi

CD30 cross-reactivity and expression in feline normal tissues and lymphomas

CD30 is a transmembrane glycoprotein of the tumor necrosis factor receptor superfamily included in the diagnostic algorithm of human cutaneous, anaplastic large cell and Hodgkin lymphomas and represents an optimal therapeutic target for CD30+ tumors. Similar diagnostic and therapeutic approaches are largely missing for feline lymphomas. Cross-reactivity of the antihuman CD30 receptor clone Ber-H2 was investigated in feline lymphomas. Comparative analysis of feline and human CD30 identified 61% identity of the amino acid sequence, with 100% identity of the main sequence of the epitope targeted by the antibody (RKQCEPDYYL). CD30 expression in normal feline tissues was restricted to rare lymphoid cells in perifollicular and interfollicular lymph node areas and in the thymic medulla. In feline lymphoma, CD30 was expressed in 4 of 33 (13%) T-cell lymphomas, 3 of 22 (14%) B-cell lymphomas, and 5 of 7 (71%) mixed-cell lymphomas, showing diffuse (1/5) or multifocal (4/5) positivity restricted to neoplastic multinucleated lymphoid cells and binucleated cells consistent with Reed-Sternberg-like cells. Based on the human classification system, cell morphology, expression of multiple markers (mixed cell components), and CD30 positivity, these cases were considered most consistent with classical Hodgkin-like lymphoma (HLL). The other 2 mixed-cell lymphomas were CD30 negative and thus most consistent with either T-cell-rich large B-cell lymphoma (TCRLBCL) or nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL). These findings provide multiple data supporting the cross-reactivity of the Ber-H2 anti-CD30 clone in feline tissues and give evidence of the usefulness of CD30 in the diagnostic evaluation of feline lymphoma

Productive results, oxidative stress and contaminant markers in european sea bass: Conventional vs. organic feeding

In the present study European sea bass (Dicentrarchus labrax) subjected to two different diets (organic vs. conventional) were evaluated in terms of growing performances, oxidative stress, and contaminant markers. Growing performances were evaluated using biometric measures and condition factor (K), whereas insulin-like growth factor (IGF-I and IGF-II) levels were assessed trough Real-Time PCR analysis. For oxidative stress, immunohistochemical staining for 8-hydroxy-2\u2032-deoxyguanosine (8-OHdG) and 4-hydroxy-2-nonenal (HNE) was performed, whereas total glutathione (GSH) in blood serum was determined by an enzymatic method adapted. Cytochrome P4501A (CYP1A) and melanomacrophage centers (MMCs) were evaluated as contaminant markers trough immunohistochemical and histochemical approaches, respectively. The growing performances showed a positive trend in both groups but a greater productivity in conventional fed fish compared to the organic ones. A significant higher expression of MMCs was observed in organic vs. conventional diet fed fish. Fillet analysis showed a higher MUFA content and a lower PUFAs n-6 content in organically fed sea bass indicating that diets with a content in fatty acids closer to that of wild fish will definitely affect the fatty acid profile of the fish flesh. On the other hand, the diet composition did not seem to affect neither the oxidative stress parameters (GSH, 8-OHdG, HNE) nor the CYP1A expression

Exposure to vector-borne pathogens in candidate blood donor and free-roaming dogs of northeast Italy.

BACKGROUND: Many vector-borne pathogens including viruses, bacteria, protozoa and nematodes occur in northeast Italy, representing a potential threat to animal and human populations. Little information is available on the circulation of the above vector-borne pathogens in dogs. This work aims to (i) assess exposure to and circulation of pathogens transmitted to dogs in northeast Italy by ticks, sandflies, and mosquitoes, and (ii) drive blood donor screening at the newly established canine blood bank of the Istituto Zooprofilattico Sperimentale delle Venezie. METHODS: Blood samples from 150 privately-owned canine candidate blood donors and 338 free-roaming dogs were screened by serology (IFA for Leishmania infantum, Ehrlichia canis, Anaplasma phagocythophilum, Babesia canis, Rickettsia conorii, R. rickettsii), microscopic blood smear examination, and blood filtration for Dirofilaria spp. All candidate donors and seropositive free-roaming dogs were tested by PCR for L. infantum, E. canis, A. phagocythophilum, Babesia/Theileria and Rickettsia spp. The dogs had no clinical signs at the time of sampling. RESULTS: Overall, 40 candidate donors (26.7 %) and 108 free-roaming dogs (32 %) were seroreactive to at least one vector-borne pathogen. Seroprevalence in candidate donors vs free-roaming dogs was: Leishmania infantum 6.7 vs 7.1 %; Anaplasma phagocytophilum 4.7 vs 3.3 %; Babesia canis 1.3 vs 2.7 %; Ehrlichia canis none vs 0.9 %; Rickettsia conorii 16 vs 21.3 % and R. rickettsii 11 vs 14.3 %. Seroreactivity to R. rickettsii, which is not reported in Italy, is likely a cross-reaction with other rickettsiae. Filariae, as Dirofilaria immitis (n = 19) and D. repens (n = 2), were identified in free-roaming dogs only. No significant differences were observed between candidate donors and free-roaming dogs either in the overall seroprevalence of vector-borne pathogens or for each individual pathogen. All PCRs and smears performed on blood were negative. CONCLUSIONS: This study demonstrated that dogs are considerably exposed to vector-borne pathogens in northeast Italy. Although the dog owners reported regularly using ectoparasiticides against fleas and ticks, their dogs had similar exposure to vector-borne pathogens as free-roaming dogs. This prompts the need to improve owner education on the use of insecticidal and repellent compounds in order to reduce the risk of arthropod bites and exposure to vector-borne pathogens. Based on the absence of pathogens circulating in the blood of healthy dogs, the risk of transmission of these pathogens by blood transfusion seems to be low, depending also on the sensitivity of the tests used for screening

DNA and RNA isolation from canine oncologic formalin-fixed, paraffin-embedded tissues for downstream "-omic" analyses: possible or not?

Formalin-fixed, paraffin-embedded (FFPE) tissues represent a unique source of archived biological material, but obtaining suitable DNA and RNA for retrospective \u201c-omic\u201d investigations is still challenging. In the current study, canine tumor FFPE blocks were used to 1) compare common commercial DNA and RNA extraction kits; 2) compare target gene expression measured in FFPE blocks and biopsies stored in a commercial storage reagent; 3) assess the impact of fixation time; and 4) perform biomolecular investigations on archival samples chosen according to formalin fixation times. Nucleic acids yield and quality were determined by spectrophotometer and capillary electrophoresis, respectively. Quantitative real-time polymerase chain reaction assays for the following genes: BCL-2\u2013associated X protein, B-cell lymphoma extra large, antigen identified by monoclonal antibody Ki-67, proto-oncogene c-KIT (c-kit). Two internal control genes (Golgin A1 and canine transmembrane BAX inhibitor motif containing 4), together with direct sequencing of c-kit exons 8, 9, 11, and 17, were used as end points. Differences in DNA/RNA yield and purity were noticed among the commercial kits. Nucleic acids (particularly RNA) extracted from paraffin blocks were degraded, even at lower fixation times. Compared to samples held in the commercial storage reagent, archived tissues showed a poorer amplification. Therefore, a gold standard protocol for DNA/RNA isolation from canine tumor FFPE blocks for molecular investigations is still troublesome. More standardized storage conditions, including time between sample acquisition and fixation, fixation time, and sample thickness, are needed to guarantee the preservation of nucleic acids and, then, their possible use in retrospective transcriptomic analysis