A Simple and Reliable Assay for Detecting Specific Nucleotide Sequences in Plants Using Optical Thin-film Biosensor Chips

Here we report the adaptation and optimization of an efficient, accurate and inexpensive assay that employs custom-designed silicon-based optical thin-film biosensor chips to detect unique transgenes in genetically modified （GM） crops and SN-P markers in model plant genomes. Briefly, aldehyde-attached sequence-specific singlestranded oligonucleotide probes are arrayed and covalently attached to a hydrazine-derivatized biosensor chip surface. Unique DNA sequences （or genes） are detected by hybridizing biotinylated PCR amplicons of the DNA sequences to probes on the chip surface. In the SN-P assay, target sequences （PCR amplicons） are hybridized in the presence of a mixture of biotinylated detector probes and a thermostable DNA ligase. Only perfect matches between the probe and target sequences, but not those with even a single nucleotide mismatch, can be covalently fixed on the chip surface. In both cases, the presence of specific target sequences is siL, nified by a color change on the chip surface （gold to blue/purple） after brief incubation with an anti-biotin IgG horseradish peroxidase （HRP） to generate a precipitable product from an HRP substrate.     

动物软组织修复资源-沙棘

伤口愈合是一种复杂的现象,是机体控制的系统对细胞进行精微同步调节的结果.然而,一系列可变因素通过多种方法来影响此过程.通常来说,机体内的各种维生素,矿物质,脂肪酸,氨基酸和其他营养物质对该过程起着直接或间接的重要影响.不仅如此,各种疾病与无数的外界因素也以不可预料的形式影响着该过程.因此,任何治疗方法都是努力控制上述因素以使机体处于最佳恢复状态.因此,当这种天然愈合过程被打断,机体的伤口就会转变为慢性,或者出现溃疡.     

Experimental infection of N'Dama cattle with trypanosomes usingGlossina palpalis gambiensis caught in the wild

Summary The transmissibility of trypanosome infection to N'Dama cattle by tsetse flies caught in the field was examined. Wild-caughtGlossina palpalis gambiensis were transferred singly into small numbered cages and allowed to feed on 14 uninfected N'Dama cattle. Following a completed feed the tsetse were dissected and infection in the proboscis, the salivary glands and the gut was recorded. Each animal was bitten by a number of tsetse ranging from five up to 64 flies. Following dissection of the tsetse flies, seven of the cattle were found to have been bitten by a single infected tsetse, five by two, while the remaining two were each fed upon by three infected tsetse. The tsetse were harbouring eitherTrypanosoma vivax or a trypanosome species belonging to theNannomonas subgenus or both species. The experimental animals were monitored daily over a period of three months for the appearance of trypanosomes in the blood and for antibodies in their sera. Other parameters such as body temperature, local skin reactions, packed red cell volume and weight changes were also measured. Trypanosomes were first detected eight days after the infective bite. Only five of the 14 cattle became infected, of which three had been exposed to a single infected tsetse fly.Trypanosoma vivax was detected in one animal,Trypanosoma congolense in two cattle and mixed infections of both species in the remaining two animals. These findings show that N'Dama cattle can become infected with trypanosomes through the bite of a single infected tsetse fly under field conditions. However, only five (possibly eight) of the 23 infected tsetse were able to transmit the parasites successfully.

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Infeccion Experimental De Ganado N'Dama Con Tripanosomas UsandoGlossina Palpalis Gambiensis Atrapadas En El Campo

Resumen Se estudió la transmisibilidad de infecciones con tripanosomas a ganado N'Dama, a través de moscas tsetse atrapadas en el campo. LasGlossina palpalis gambiensis atrapadas, fueron transferidas individualmente a jaulas numeradas, donde se alimentaron sobre 14 animales N'Dama sin infección alguna.Una vez completada una comida, las moscas se disecaron buscándose infecciones en la proboscis, glándulas salivares e intestino. Cada animal fue picado por un buen número de moscas, en un rango de cinco a 64. Seguida a la disección de las moscas tsetse, siete bovinos fueron picados por una sola mosca infectada; cinco por dos, mientras que los otros dos fueron cada uno picado por tres tsetse infectadas. Las moscas estaban infectadas conTrypanosoma vivax o con especies de tripanosomas pertenecientes al subgéneroNannomonas, o por ambas especies.Los animales fueron seguidos clínicamente día a día por un período de tres meses por la presencia de tripanosomas en la sangre y anticuerpos en el suero. Se tomaron en cuenta otros parámetros como la temperatura corporal, reacciones cutáneas locales, volumen corpuscular celular y peso corporal. Los primeros tripanosomas se detectaron ocho días después de la picadura de moscas infectadas. Solamente cinco de los 14 bovinos se infectaron, de los cuales tres, habían sido expuestos a una única tsetse infectada. Se detectóTrypanosoma vivax en un animal,T. congolense en dos bovinos más e infecciones mixtas en los otros dos.Estos hallazgos demuestran, que el ganado N'Dama puede infectarse con tripanosomiasis, seguidamente de la picadura de una sola tsetse infectada bajo condiciones de campo. Sin embargo, solamente cinco (posiblemente ocho) de las 23 tsetse infectadas, fueron capaces de transmitir el parásito exitosamente.

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Infection Experimentale De Bovins N'Dama Avec Des Trypanosomes A l'Aide DeGlossina Palpalis Gambiensis Capturees A l'Etat Sauvage

Résumé La transmissibilité de possibilité de transmission de la trypanosomose à des bovins N'Dama par des mouches tsé-tsé capturées sur le terrain a été examinée. DesGlossina palpalis gambiensis capturées à l'état sauvage ont été transférées individuellement dans de petities cages numérotées et placées pour se nourrir sur 14 bovins N'Dama indemnes.A la suite d'un repas complet, les mouches ont été disséquées et on a enregistré l'infection dans le proboscis, les glandes salivaires et l'intestin. Chaque animal a été piqué par un nombre de mouches allant de cinq à soixante quatre. A la suite de la dissection des glossines, on a trouvé que sept bovins avaient été piqués par une seule mouche infectée, cinq par deux mouches, alors que les deux derniers avaient servi à nourrir chacun trois glossines infectées. Les mouches tsé-tsé hébergeaient soitTrypanosoma vivax ou une espèce de trypanosomes appartenant au sous genreNannomonas ou par les deux espèces.Les animaux d'expérience ont été suivis quotidiennement pendant trois mois pour observer l'apparition des trypanosomes dans le sang et des anticorps dans leur sérum. D'autre paramètres tels que la température corporelle, les réactions cutanées locales, le volume erythrocytaire et les variations pondérales ont également été mesurés. Les trypanosomes ont été détectés pour la première fois huit jours après la morsure infectante. Seuls cinq des 14 bovins se sont infectés, parmi lesquels trois avaient été exposés à une seule mouche infectée. On a détéctéTrypanosoma vivax chez un animal,T. congolense chez deux bovins et des infections mixtes des deux espèces chez les deux derniers animaux infectés. Ces observations montrent que les bovins N'Dama peuvent devenir infectés par des trypanosomes par la piqure d'une unique mouche tsé-tsé infectée dans les conditions du terrain. Cependant, seulement cinq (peut-être huit) des 23 glosssines infectées ont été capables de transmettre les parasites avec succès.

     

Plasmid-encoded antibiotic resistance in Staphylococcus hyicus

A total of 33 Staphylococcus hyicus-cultures from piglets with exudative epidermatitis were analyzed for the presence of antibiotic resistance plasmids. Four small plasmids encoding resistances to chloramphenicol, macrolide-lincosamide-antibiotics, streptomycin or tetracyclines could be identified in plasmid-curing and plasmid-transformation experiments. For further characterization these plasmids were digested with restriction endonucleases. This led to the construction of a specific restriction map for each of the 4 plasmids. On the basis of their restriction maps, these 4 antibiotic resistance plasmids from "porcine" S. hyicus-cultures were compared with the respective resistance plasmids of other staphylococcal species from infections of humans and animals.     

Efficacy of ceftiofur hydrochloride for treatment of experimentally induced colibacillosis in neonatal swine

Ceftiofur hydrochloride was tested for effectiveness against induced colibacillosis in neonatal swine. In this model, pigs less than 12 hours old were inoculated via stomach tube with a virulent, K99+, nalidixic acid-resistant strain of Escherichia coli. Six hours after challenge exposure, 1 dose of ceftiofur was administered either IM or orally in experiment 1 and orally only in experiment 2. Mortality, shedding of bacteria, fecal consistency scores, and body weight changes were monitored for 10 days. In experiment 1 (n = 383 pigs), all treatments at dosage that ranged between 0.5 and 64.0 mg of ceftiofur/kg of body weight significantly (P less than 0.001) reduced mortality, bacterial shedding, and diarrhea and increased weight gain, compared with findings in untreated controls. There were no detectable differences between oral and IM routes, except that there was greater reduction in bacteria shedding associated with the oral route of administration. In experiment 2 (n = 505 pigs), ceftiofur was administered orally either once at 6 hours after challenge exposure or twice at 6 and at 48 hours after the first dose. Dosage of ceftiofur was 0, 5, 10, 20, 30, or 60 mg/kg administered once, or half the same dose was administered at each of 2 times. At the optimal dosage (10 mg/kg), a single dose was as effective as 2 doses. The single administration at all dosages reduced mortality, bacterial shedding, and diarrhea scores and increased body weight gain, compared with findings in untreated pigs (P less than 0.01). In this induced infection model, the optimal treatment dosage was determined to be 10 mg/kg administered once.     

Plasma von Willebrand factor concentration and thyroid function in dogs

Plasma von Willebrand factor antigen concentration was determined in 15 dogs with suspected hypothyroidism, in 1 dog with hyperthyroidism, and in 14 euthyroid dogs. The mean +/- SEM von Willebrand factor:antigen concentration in hypothyroid dogs (47.1% +/- 12.6%) was significantly decreased (P less than 0.0005), compared with that in euthyroid dogs (94.7 +/- 5.6%). Four hypothyroid dogs were given thyroxine for 1 month and all 4 had an increase in von Willebrand factor:antigen concentration. The plasma von Willebrand factor:antigen concentration was 200% in the hyperthyroid dog. Seemingly, reduced concentrations of plasma von Willebrand factor:antigen can be found in dogs in association with congenital von Willebrand disease or with von Willebrand disease acquired through hypothyroidism.     

Pathogenicity of peste des petits ruminants virus isolated from Egyptian goats in Egypt

2 Egyptian goats and Boscat rabbits were experimentally inoculated with peste des petits ruminants (PPR) local Egyptian strain (PPR, Egypt 87). The inoculated animals contracted the disease with minor clinical manifestations, accompanied by rise of neutralizing antibodies to PPR virus. Virus was isolated from ocular and nasal secretions, buffy coat, spleen, and liver. No contact infection was observed between inoculated and healthy goats.     

Enteral nutrition by tube

When oral intake is unsatisfactory or contraindicated, maintenance of nutrition by tube feeding is an alternative to the parenteral route. A large volume of research data supports the decision to use the enteral route whenever possible. Entry of food into the alimentary tract is a stimulus to structural and functional maintenance of that tract. Enteral nutrition can be given via indwelling nasoesophageal, pharyngostomy, esophagostomy, percutaneous or surgical gastrostomy, or enterostomy tube. Use of an appropriate catheter, familiarity with the technique used, and careful patient selection and monitoring are important factors in successful tube feeding. Blenderized pet food diets should be fed whenever possible; commercially available liquid diets provide an alternative when tube caliber or patient factors preclude the use of blenderized foods.     

Use of ELISA for detection of immunoglobulins G and M that recognize Salmonella dublin lipopolysaccharide for prediction of carrier status in cattle

Immunoglobulin reactions to Salmonella dublin in serum and milk from 4 groups of lactating cows were measured by an indirect ELISA. The groups consisted of (1) cows that were natural carriers of S dublin in the mammary gland, (2) experimentally infected cows that did not become carriers, (3) cows inoculated with a commercial S dublin bacterin, and (4) cows used as S dublin-negative controls. Milk and serum samples were obtained at monthly intervals. Models for predicting carrier status were developed by use of stepwise logistic regression. Independent variables consisted of serum and milk IgG and IgM titers to S dublin lipopolysaccharide and a ratio of IgG to IgM. The utility of a single sample vs multiple samples obtained at 1-month or 2-month intervals was tested by comparison of goodness-of-fit chi 2 P values for 8 models predicting carrier status. Immunoglobulin reactions specific to S dublin were a significant predictor of carrier status (P less than 0.001). Serum IgG titers specific for S dublin were the most important variable for predicting carrier status. Two serum IgG titers to S dublin obtained 2 months apart was a better predictor of carrier status than measurement of the IgG:IgM ratio from a single serum sample. Immunoglobulin recognizing S dublin epitopes also were detected in milk samples. In milk, performing 2 ELISA 60 days apart to determine IgG and IgM reactions to S dublin appeared to be useful for the prediction of carrier status, but was not as accurate as models for serum immunoglobulin reactions.     

Procollagen type-III aminoterminal peptide in serum and synovial fluid of dogs with hip dysplasia and coxarthrosis

Hip dysplasia is an affection of the coxofemoral joint that progresses until stabilization is caused by fibrosis and osteoarthritic changes. This stabilization process can be examined by clinical and radiographic methods. The capability of evaluating the procollagen concentrations in liquids, such as serum and synovial fluid, has further offered the basis for an objective biochemical evaluation of the stabilization process. Our study was performed to evaluate whether determination of procollagen concentrations was suitable for the use in practice. The procollagen type-III aminoterminal peptide (P-III-NP) concentration was measured in serum and in synovial fluid from coxofemoral joints in 20 dogs. Dogs were grouped on the basis of evidence of dysplasia and osteoarthritic changes of the hip: (1) a control group of 6 dogs without clinical or radiographic signs of hip dysplasia, and (2) dysplastic group of 14 dogs, which was further grouped with respect to the coxofemoral joint laxity, as determined by the Ortolani test. Synovial fluid concentration of P-III-NP was significantly (P less than 0.05) higher in fluid from dysplastic joints than in fluid from normal joints. Serum concentrations of P-III-NP were significantly (P less than 0.05) higher in dogs in which results of the Ortolani test were positive.