Development of a genetic manipulation system for Haemophilus parasuis

Haemophilus parasuis is a member of the family Pasteurellaceae and an important respiratory-tract pathogen of swine, which is the etiological agent of Glasser's disease. Because no genetic manipulation system is available for H. parasuis so far, in vivo studies about the role of its genes involved in virulence are unfeasible. Here we demonstrate that H. parasuis has a cyclic AMP (cAMP)-dependent natural transformation system that enables the uptake of DNA in which the ACCGAACTC sequence signal must be present. After improving DNA transformation parameters, such as cAMP and DNA concentration and exposition time of the exogenous DNA, a knockout mutant of H. parasuis defective in the thy gene, encoding the thymidylate synthase enzyme, has been constructed. Data presented in this work open the possibility for the functional analysis of genes involved in the infectious process of this animal pathogen.     

The prevalence of Toxocara cati in domestic cats in Mexico City

The faeces of 520 domestic cats, resident in the 16 municipal authorities of Mexico City, were analysed using the method of centrifuging with zinc sulphate (at 33%). Three hundred and ninety-nine animals lived in houses, 121 in apartments. For the purpose of this study they were divided into seven age groups, of 6 months each. Toxocara cati eggs were found in the faeces of 42.5% of the animals. 20.7% of apartment cats and 49.1% of house cats were infected. This difference in the rate of infection is statistically significant (P<0.002.) for the house cats. T. cati was found in all the age groups although most infection was found in animals less than 1 year old.The fact that infection occurred in the whole age range of the cats and in all the municipal authorities means that the risk of being contaminated with this helminth in Mexico City is high. As a consequence, the possibility of developing illness as a result of becoming a host to larva migrans is also high.     

Design, validation, and absolute sensitivity of a novel test for the molecular detection of avian pneumovirus.

This study describes attempts to increase and measure sensitivity of molecular tests to detect avian pneumovirus (APV). Polymerase chain reaction (PCR) diagnostic tests were designed for the detection of nucleic acid from an A-type APV genome. The objective was selection of PCR oligonucleotide combinations, which would provide the greatest test sensitivity and thereby enable optimal detection when used for later testing of field materials. Relative and absolute test sensitivities could be determined because of laboratory access to known quantities of purified full-length DNA copies of APV genome derived from the same A-type virus. Four new nested PCR tests were designed in the fusion (F) protein (2 tests), small hydrophobic (SH) protein (1 test), and nucleocapsid (N) protein (1 test) genes and compared with an established test in the attachment (G) protein gene. Known amounts of full-length APV genome were serially diluted 10-fold, and these dilutions were used as templates for the different tests. Sensitivities were found to differ between the tests, the most sensitive being the established G test, which proved able to detect 6,000 copies of the G gene. The G test contained predominantly pyrimidine residues at its 3' termini, and because of this, oligonucleotides for the most sensitive F test were modified to incorporate the same residue types at their 3' termini. This was found to increase sensitivity, so that after full 3' pyrimidine substitutions, the F test became able to detect 600 copies of the F gene.     

Intra-abdominal leiomyosarcoma in a ferret (Mustela putorius Furo): histopathological and immunohistochemical characterization

A 5 years old female ferret with an abdominal palpable mass confirmed at echo-graphic examination died during an explorative laparotomy. A single lymph-node-like nodule was found adjacent to the intestinal loops. The round mass well circumscribed, solid and white, histologically, at low magnification, appeared encapsulated and built up by a population of atypical spindle cells arranged in interwoven fascicles. The cells had high anisocytosis degree, moderate mitotic activity and prominent nucleoli. A central area of necrosis was present. To characterise the tumour immunohistochemically cytokeratin, vimentin, S-100, melan-A, vWF, desmin, actin and alpha-actin were applied. Neoplastic cells resulted positive to vimentin, actin and alpha-actin. Based on the histological and immunohistological pattern a diagnosis of leiomyosarcoma was made.     

Echinochrome A Treatment Alleviates Atopic Dermatitis-like Skin Lesions in NC/Nga Mice via IL-4 and IL-13 Suppression

Atopic dermatitis (AD) is a chronic inflammatory skin disease in which skin barrier dysfunction leads to dryness, pruritus, and erythematous lesions. AD is triggered by immune imbalance and oxidative stress. Echinochrome A (Ech A), a natural pigment isolated from sea urchins, exerts antioxidant and beneficial effects in various inflammatory disease models. In the present study, we tested whether Ech A treatment alleviated AD-like skin lesions. We examined the anti-inflammatory effect of Ech A on 2,4-dinitrochlorobenzene (DNCB)-induced AD-like lesions in an NC/Nga mouse model. AD-like skin symptoms were induced by treatment with 1% DNCB for 1 week and 0.4% DNCB for 5 weeks in NC/Nga mice. The results showed that Ech A alleviated AD clinical symptoms, such as edema, erythema, and dryness. Treatment with Ech A induced the recovery of epidermis skin lesions as observed histologically. Tewameter^® and Corneometer^® measurements indicated that Ech A treatment reduced transepidermal water loss and improved stratum corneum hydration, respectively. Ech A treatment also inhibited inflammatory-response-induced mast cell infiltration in AD-like skin lesions and suppressed the expression of proinflammatory cytokines, such as interferon-γ, interleukin-4, and interleukin-13. Collectively, these results suggest that Ech A may be beneficial for treating AD owing to its anti-inflammatory effects.     

Mapping sub-field maize yields in Nebraska,USA by combining remote sensing imagery,crop simulation models,and machine learning

Precision Agriculture - Crop yield maps are valuable for many applications in precision agriculture, but are often inaccessible to growers and researchers wishing to better understand yield...     

Measurement of progesterone in sheep using a commercial ELISA kit for human plasma

Determination of serum or plasma progesterone (P4) concentrations is important to recognize pregnant and non-pregnant ewes, and also to predict the number of carried lambs. The 2 most common methodologies for the detection of plasma P4 are radioimmunoassay (RIA) and enzyme immunoassay (EIA). RIA is very expensive, and not all laboratories are equipped to perform this test; EIA is commercially available for human use, but only a few companies produce species-specific kits, which are expensive. We verified for ovine plasma a less expensive and easily available ELISA kit (DiaMetra) designed to quantify P4 in humans. Pools of ovine and human plasma were used to compare repeatability, accuracy, sensitivity, and stability of P4 measured by the DiaMetra kit. Repeatability data were within 15%, and accuracy values were ~90% for both plasma matrices. Stability data showed a loss of <20% for freeze–thaw and <30% for 30-d storage. All parameters were acceptable under international guidelines for method validation. The human ELISA kit was used successfully to quantify plasma P4 in 26 ewes during pregnancy until delivery. P4 concentrations were also correlated with the number of carried lambs.     

Non‐indigenous fish in protected spaces: Trends in species distribution mediated by illegal stocking

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