A rapid multiplexed chemiluminescent immunoassay for the detection of Escherichia coli O157:H7, Yersinia enterocolitica, Salmonella typhimurium, and Listeria monocytogenes pathogen bacteria

A simple and rapid multiplexed sandwich chemiluminescent enzyme immunoassay has been developed for the simultaneous detection of Escherichia coli O157:H7, Yersinia enterocolitica, Salmonella typhimurium, and Listeria monocytogenes. To achieve the multiplexed detection of the four pathogens, a new polystyrene 96 well microtiter plate format has been designed, in which each main well contains four subwells in the bottom. The monoclonal antibodies specific for each bacteria were separately immobilized in each subwell. When the samples were added to the main wells, the bacteria able to specifically bind to the corresponding monoclonal antibody were captured in one of the four subwells. Subsequently, a mixture of peroxidase-labeled polyclonal antibodies against the four bacteria was added and the peroxidase activity of the bound polyclonal labeled antibodies in each well was measured by an enhanced luminol-based chemiluminescent cocktail using a low-light charge-coupled imaging device. The assay was simple and fast, and the limit of quantification was in the order of 104-105 CFU/mL for all bacterial species. The accuracy of the method, evaluated by comparison of the results with a conventional culturing methodology, was satisfactory, with recovery values ranging from 90 to 120%. This method can be used as a screening test to evaluate the presence of these pathogen bacteria in different foodstuffs.     

Antioxidant activity of Sicilian pistachio (Pistacia vera L. var. Bronte) nut extract and its bioactive components

Pistacia vera L. is the only species of Pistacia genus producing edible nuts. This paper investigates the antioxidant potential of a Sicilian variety of pistachio nut by chemical as well as biological assays and measured antioxidant vitamins and a number of antioxidant polyphenols in either the hydrophilic and/or the lipophilic nut extract. In accordance with the majority of foods, the total antioxidant activity, measured as a TAA test, was much higher (50-fold) in the hydrophilic than in the lipophilic extract. Substantial amounts of total phenols were measured. The hydrophilic extract inhibited dose-dependently both the metal-dependent and -independent lipid oxidation of bovine liver microsomes, and the Cu+2-induced oxidation of human low-density lipoprotein (LDL). Peroxyl radical-scavenging as well as chelating activity of nut components may be suggested to explain the observed inhibition patterns. Among tocopherols, gamma-tocopherol was the only vitamin E isomer found in the lipophilic extract that did not contain any carotenoid. Vitamin C was found only in a modest amount. The hydrophilic extract was a source of polyphenol compounds among which trans-resveratrol, proanthocyanidins, and a remarkable amount of the isoflavones daidzein and genistein, 3.68 and 3.40 mg per 100 g of edible nut, respectively, were evaluated. With the exception of isoflavones that appeared unmodified, the amounts of other bioactive molecules were remarkably reduced in the pistachio nut after roasting, and the total antioxidant activity decreased by about 60%. Collectively, our findings provide evidence that the Sicilian pistachio nut may be considered for its bioactive components and can effectively contribute to a healthy status.     

A convenient route to the synthesis of isotopomeric dihydro-2(3H)furanones

A general synthetic procedure leading to isotopomeric dihydro-2(3H)furanones (gamma-butyrolactones) containing two, four, or six deuterium atoms has been developed. The labeled dihydro-2(3H)furanones were synthesized in quantitative yield from the saturated diacid C4 (succinic) or unsaturated diacids C4 (fumaric, maleic, or acetylendicarboxylic) in the presence of Ru4H4(CO)8(PBu3)4 using a deuterium pressure of 180 bar at 180 degrees C. This methodology was applied to the total synthesis of a hexadeuterated matairesinol lignan: The 3,4-bis[[3-methoxy-4-(phenylmethoxy)phenyl]methyl]dihydro-2(3H)furanone-[7,7',8,8',9',9'-D6] (benzyl-protected matairesinol-D6) was fully characterized.     

Endogenous retroviruses of sheep: a model system for understanding physiological adaptation to an evolving ruminant genome

Endogenous retroviruses (ERVs) are present in the genome of all vertebrates and are remnants of ancient exogenous retroviral infections of the host germline transmitted vertically from generation to generation. Sheep betaretroviruses offer a unique model system to study the complex interaction between retroviruses and their host. The sheep genome contains 27 endogenous betaretroviruses (enJSRVs) related to the exogenous and pathogenic Jaagsiekte sheep retrovirus (JSRV), the causative agent of a transmissible lung cancer in sheep. The enJSRVs can protect their host against JSRV infection by blocking early and late steps of the JSRV replication cycle. In the female reproductive tract, enJSRVs are specifically expressed in the uterine luminal and glandular epithelia as well as in the conceptus (embryo and associated extraembryonic membranes) trophectoderm and in utero loss-of-function experiments found the enJSRVs envelope (env) to be essential for conceptus elongation and trophectoderm growth and development. Collectively, available evidence in sheep and other mammals indicate that ERVs coevolved with their hosts for millions of years and were positively selected for biological roles in genome plasticity and evolution, protection of the host against infection of related pathogenic and exogenous retroviruses, and placental development.     

Stomach wall evaluation using helical hydro-computed tomography

In helical hydro-computed tomography (helical hydro-CT), water is used as a neutral luminal contrast medium together with intravenous iodine contrast medium for the diagnosis and staging of human gastric neoplasia. We evaluated the feasibility of helical hydro-CT in 11 healthy animals (nine dogs and two cats). Adequate uniform gastric distension was obtained with 30 ml water/kg body weight. Fourteen client-owned dogs and four cats with suspected or diagnosed gastric neoplasia then underwent helical hydro-CT followed by intravenous contrast medium administration. Focal thickening with moderate contrast enhancement was found in 10 dogs and 3 cats. The extent of the lesion was assessed easily in all these patients. Three dogs and one cat had a normal stomach wall. One dog had multifocal thickening of the antrum but no histopathologic diagnosis was made. Helical hydro-CT, followed by intravenous contrast medium administration, is a simple technique for assessing the stomach wall.     

Laparoscopic sterilization of Sardinia donkeys using an endoscopic stapler

OBJECTIVE: To describe a laparoscopic technique for sterilization of the male donkey using an endoscopic linear stapler (ELS). STUDY DESIGN: Clinical report. ANIMALS: Male Sardinia donkeys (n=6), aged 24-36 months. METHODS: Laparoscopic sterilization was performed under standing sedation or recumbent under general anesthesia. An endoscopic linear stapler was used to transect, and control hemorrhage of, the spermatic cord. Serum testosterone concentrations were measured preoperatively and at 3, 6, and 12 months after sterilization. One testis from each of 2 donkeys was collected at 12 months for histologic examination. RESULTS: Sterilization was successfully achieved. The only minor intraoperative complication experienced was hemorrhage from 1 spermatic cord stump, which was re-stapled. No other major short- or long-term complications occurred. Serum testosterone concentrations were < 15 ng/dL at 3, 6, and 12 months after surgery in all donkeys. Testicular atrophy without evidence of revascularization was observed in the 2 testes examined histologically. CONCLUSIONS: Sterilization of descended testes by intraabdominal laparoscopic transection of the spermatic cord is a simple practical method, with minimal intraoperative or postoperative complications. Testicular revascularization did not occur. CLINICAL RELEVANCE: Laparoscopic sterilization in donkey can be performed using an endoscopic linear stapler as an alternative to scrotal castration.     

Computed tomography-guided fine-needle aspiration and tissue-core biopsy of bone lesions in small animals

In humans, free-hand computed tomography (CT)-guided biopsy is an accurate method to obtain a tissue sample. There are only a few reports of this technique in veterinary medicine. In the present study, 21 dogs and two cats underwent a free-hand CT-guided tissue-core biopsy (17 animals) or fine-needle aspiration (six animals) of a bone lesion. Two out of 17 tissue-core samples were also cultured. All 17 tissue-core biopsy samples were diagnostic (accuracy of 100%). Five out of six aspirates were diagnostic (accuracy of 83.3%). The overall accuracy was 95.7%. In one aspirate, cytologic quality was insufficient containing only blood. No major complications were encountered. Fourteen neoplastic, two infectious and six benign lesions were diagnosed. CT examination after intravenous contrast medium added useful information to avoid large vessels and to biopsy-viable tissue. Free-hand CT-guided tissue-core biopsy and aspiration appears to be a safe and very accurate procedure for use in the diagnosis of bone-associated diseases in small animals.     

Fatal necrotizing fasciitis and myositis in a captive common bottlenose dolphin (Tursiops truncatus) associated with Streptococcus agalactiae.

A common bottlenose dolphin (Tursiops truncatus) was presented for necropsy after acute onset of gastrointestinal signs and cutaneous lesions that rapidly progressed to death. Gross and microscopic findings were characterized by locally extensive severe necrohemorrhagic fasciitis and cellulitis, and severe necrotizing myositis in the head and dorsocranial thorax, with numerous disseminated gram-positive cocci. Streptococcus agalactiae was isolated from the lesions and from visceral organs (liver and lung), and it was identified by standard microbiology techniques. This communication is the first report of necrotizing fasciitis in a marine mammal associated with S. agalactiae.     

SPINAL SUBARACHNOID CYST IN A CAT

A 7-year-old neutered female domestic short hair cat was presented with hind limb ataxia. A subarachnoid cyst in the T10-T11 spinal cord region was identified by myelography as a collection of contrast medium in the subarachnoid space.     

Effects of compost and defatted oilseed meals as sustainable organic fertilisers on cardoon (Cynara cardunculus L.) production in the Mediterranean basin

Cardoon (Cynara cardunculus L.) is considered as one of the most suitable energy crops for Southern Europe. The aim of this work was to outline the effects of organic fertilisers on the productivity and the global warming potential (GWP) on cardoon production. Six fertilisers (N 100 kg ha^?1, N 50 kg ha^?1, Compost 30 t ha^?1, Compost 15 t ha^?1 + N 25 kg ha^?1, 3 t ha^?1 of defatted oilseed meals of sunflower, 3 t ha^?1 of defatted oilseed meals of Brassica carinata), and unfertilised control, were evaluated on two cultivars (‘Gobbo di Nizza’ and ‘Altilis 41’) in a split-plot experiment. Defatted oilseed meal of sunflower recorded higher total dry weight (+10%), seed yield (+17%), nitrogen use efficiency (+14%) and better GWP (?66%) compared to the other organic fertilisers and performing as well as N 100 kg ha^?1. Altilis 41 cultivar showed the highest aboveground total dry weight (10 t ha^?1 y^?1), seed yield (1.7 t ha^?1 y^?1), stalk dry weight (7 t ha^?1 y^?1) and head dry weight (3 t ha^?1 y^?1). Our results highlighted that by combining suitable cultivar and fertilisation strategies, it could be possible to increase the production sustainability of C. cardunculus.