Rapid detection of Salmonella from poultry by real-time polymerase chain reaction with fluorescent hybridization probes

Detection of Salmonella by bacteriologic methods is known to be time consuming. Therefore, we have developed a real-time probe-specific polymerase chain reaction (PCR) to rapidly detect Salmonella invA gene-based PCR products from chicken feces and carcasses by a fluorescence resonance energy transfer assay. The sensitivity and the specificity of this system were determined as 3 colony-forming units ml(-1) and 100%, respectively. Overnight tetrathionate broth enrichment cultures of chicken feces and carcass samples were used in template preparation for PCR. Also, a standard bacteriology was performed (National Poultry Improvement Plan-U.S. Department of Agriculture, Bacteriological Analytical Manual-Food and Drug Administration Center for Food Safety and Applied Nutrition) for confirmation. Seventy-two cloacal swab, 147 intestine, and 50 carcass (neck) samples were examined. Thirteen (8.8%) and 25 (17%) of the intestinal samples were found to harbor Salmonella by bacteriology and PCR, respectively. Forty-five of 50 (90%) carcass samples were Salmonella positive by both methods. Salmonella was not detected from cloacal swab samples. Results indicate that this assay has the potential for use in routine monitoring and detection of Salmonella in infected flocks and carcasses.     

Pharmacokinetics and perioperative efficacy of intravenous ketorolac in dogs

Ketorolac (KET) is a nonsteroidal anti‐inflammatory drug approved for the use in humans that possesses a potent analgesic activity, comparable to morphine, and could represent a useful tool to control acute pain also in animals. The clinical efficacy and pharmacokinetic profile of intravenous (IV) ketorolac tromethamine (0.5 mg/kg) were studied in 15 dogs undergoing gonadectomy. Intra‐operative cardiorespiratory variables were monitored, and post‐operative pain was assessed using a subjective pain score (0–24) in all dogs, whereas the pharmacokinetic profile of the drug was determined in 10 animals. During surgery, mean minimal alveolar concentration of isoflurane was 1.69 ± 0.11%, and normocapnia and spontaneous ventilation were maintained in all animals. During pain assessment, no significant differences between males and females were found, and in no case rescue analgesia was necessary. No adverse effects were reported. Serum samples were purified by solid‐phase extraction and analysed by HPLC with UV‐Vis detection. A large variability was observed in serum concentrations. The kinetics of ketorolac was described by a noncompartmental analysis. The elimination half‐life (t_½λz) and Cl_B were 10.95 ± 7.06 h and 92.66 ± 84.49 mL/h/kg, respectively, and V_dss and V_z were 1030.09 ± 620.50 mL/kg and 1512.25 ± 799.13 mL/kg, respectively. AUC_(0→last) and MRT_(0→last) were 6.08 ± 3.28 h × μg/mL and 5.59 ± 2.12 h, respectively. The results indicate that ketorolac possess good post‐operative analgesic effects until about 6 h after administration in dogs undergoing moderately painful surgery.     

Clinicopathological findings, molecular detection and characterization of Babesia gibsoni infection in a sick dog from Italy

A 4-year-old intact female American Pit Bull Terrier from Italy descendant of an American-born bitch was evaluated for anorexia, lethargy, weakness, and intermittent vomiting. On physical examination, the dog was dehydrated, had pale mucous membranes, hunched posture and abdominal pain. A moderate anemia was observed. Splenomegaly and hyperechoic regions suspected as infarcts in the spleen were seen on abdominal ultrasound. Based on the suspicion of splenic torsion, splenectomy was performed. After surgery, the clinical condition deteriorated. A follow-up complete blood count demonstrated severe macrocytic normochromic anemia with evidence of marked regeneration, left shift neutrophilia, monocytosis and marked thrombocytopenia. Blood smear evaluation revealed single to multiple, variable sized (1–3 μm in diameter), and round to oval to band-like piroplasms within many red blood cells consistent with small form Babesia spp. or Theileria spp. A partial segment of the 18S rRNA gene was amplified and the PCR product was analyzed by direct sequencing. The nucleotide sequence was completely identical to that of Babesia gibsoni present in GenBank^®. This is the first molecular detection and characterization of B. gibsoni infection in a sick dog from Italy.     

Absorption kinetics and bioavailability of cephalexin in the dog after oral and intramuscular administration

The pharmacokinetics of cephalexin, a first generation cephalosporin, were investigated in dogs using two formulations marketed for humans, but also often employed by practitioners for pet therapy. Cephalexin was administered to five dogs intravenously and intramuscularly as a sodium salt and by the oral route as a monohydrate. The dosage was always 20 mg/kg of active ingredient. A microbiological assay with Sarcina lutea as the test organism was adopted to measure cephalexin concentrations in serum. The mean residence time (MRT) median values after intravenous (i.v.), intramuscular (i.m.) and oral administration (p.o.) were 86 min, 200 min, and 279 min, respectively. After i.m. and oral dosing the peak serum concentrations (24.2 +/- 1.8 micrograms/mL and 20.3 +/- 1.7 micrograms/mL, respectively) were attained at 90 min in all dogs and bioavailabilities were 63 +/- 10% and 57 +/- 5%, respectively. The time course of the cephalexin serum concentrations after oral administration was best described by a model incorporating saturable absorption kinetics of the Michaelis-Menten type: thus in the gastrointestinal tract of dogs a carrier mediated transport for cephalexin similar to that reported in humans, may exist. The predicted average serum concentrations of cephalexin after repeated i.m. and oral administration indicated that, in order to maintain the therapeutic concentrations, the 20 mg/kg b.w. dosage should be administered every 6-8 h.     

Potato <Emphasis Type="Italic">R1</Emphasis> resistance gene confers resistance against <Emphasis Type="Italic">Phytophthora infestans</Emphasis> in transgenic tomato plants

Tomato is challenged by several pathogens which cause loss of production. One such pathogen is the oomycete Phytophthora infestans which is able to attack all the aerial parts of the plant. Although a wide range of resistance sources are available, genetic control of this disease is not yet successful. Pyramiding R-genes through genetic transformation could be a straightforward way to produce tomato and potato lines carrying durable resistance to P. infestans. In this work the R1 potato gene was transferred into tomato lines. The tomato transgenic lines were analyzed by using q-RT-PCR and progeny segregation to determine the gene copy number. To test the hypothesis that R1 represents a specifically regulated R-gene, transgenic tomato plants were inoculated with P. infestans isolate 88133 and IPO. All the plants containing the R1 gene were resistant to the late blight isolate IPO-0 and susceptible to isolate 88133. These results provide evidence for specific activation of the R1 gene during pathogen challenge. Furthermore, evidence for enhancement of PR-1 gene expression during P. infestans resistance response was obtained.     

Pharmacokinetics, intraoperative effect and postoperative analgesia of tramadol in cats

Tramadol is a synthetic codeine analogue used as an analgesic in human and veterinary medicine, but not approved for use in cats. Tramadol (2 mg/kg) was administered intravenously (IV) as preoperative analgesic in 12 cats (6 males) undergoing surgical gonadectomy. The pharmacokinetic profile of the drug and its O-desmethyl metabolite were determined in 8 animals (4 males), while intraoperative effects and postoperative analgesia, estimated by subjective pain score (0–24), were evaluated in all. Mean intraoperative isoflurane consumption was reduced, but hypoventilation was not observed. Sex-related differences were not observed, particularly in terms of postoperative analgesia: rescue analgesic was never administered. Concentrations of the active O-desmethyl metabolite were persistently high in all the animals. Considering the results obtained in this study, tramadol, at the dose of 2 mg/kg IV, did not produce any evident intraoperative cardiorespiratory side effects and with additional investigation may prove to be an appropriate intraoperative analgesic in cats undergoing gonadectomy.     

Maskelynite: Formation by Explosive Shock

When high pressure (250 to 300 kilobars) was applied suddenly (shock-loading) to gabbro, the plagioclase was transformed to a noncrystalline phase (maskelynite) by a solid-state reaction at a low temperature, while the proxene remained crystalline. The shock-loaded gabbro resembles meteorites of the shergottite class; this suggests that the latter formed as a result of shock. The shock-loading of gabbro at 600 to 800 kilobars raised the temperature above the melting range of the plagioclase.     

Physiological responses of Quercus ilex Leaves to Water Stress and Acute Ozone Exposure Under Controlled Conditions

The combined effect of water stress and ozone (O₃) on stomatal O₃ flux, damage to photosynthesis, and detoxification by biogenic volatile organic compounds (BVOC) in Quercus ilex leaves was studied. A 4-weeks O₃ exposure (250 ppb, 4 h per day) caused a reduction of photosynthesis and stomatal conductance, which was fully recovered 1 week after the end of the treatment, in well-watered and water-stressed plants. Measurements of stomatal O₃ flux revealed a low stomatal flux of the pollutant, which became minimal after stomatal closure caused by water stress. An induction of volatile monoterpenes, important compounds in the O₃ scavenging system in Q. ilex, and a burst of lipoxygenase compounds (LOX), which are released as gaseous by-products of membrane peroxidation, was observed after 2–3 weeks of O₃ fumigation. However, these compounds were also released in control leaves that were exposed to ozone only briefly, to determine stomatal O₃ flux. The low stomatal flux that occurred in water stress conditions helped avoiding permanent damage to Q. ilex leaves, although during the O₃ treatment photosynthesis was severely limited by stomatal closure. In well-watered plants, O₃ fumigation caused a noticeable increase of nocturnal stomatal conductance. If confirmed on adult plants under field conditions, this effect can imply larger flux of O₃ at night and possible detrimental effects of O₃ on leaf functions in plants exposed to high nocturnal O₃ levels.     

Contributions of ethnobiology to the conservation of tropical rivers and streams

• 1. This study aimed to link basic ethnobiological research on local ecological knowledge (LEK) to the conservation of Brazilian streams, based on two case studies: original data on LEK of fishermen about freshwater fish in the Negro River, Amazon, and previously published data about LEK of farmers on the ecological relationship between forest and streams in the Macabuzinho catchment, Atlantic Forest.
• 2. Information was obtained from fishermen through interviews using standard questionnaires containing open-ended questions. Informants for interview were selected either following some defined criteria or applying the ‘snowball’ method.
• 3. Fishermen's LEK about the diets and habitats of 14 fish species in the Negro River provided new biological information on plant species that are eaten by fish, in addition to confirming some ecological patterns from the biological literature, such as dependence of fish on forests as food sources.
• 4. In the Atlantic Forest, a comparison between farmers' LEK and a rapid stream assessment in the farmers' properties indicated that farmers tended to overestimate the ecological integrity of their streams. Farmers recognized at least 11 forest attributes that correspond to the scientific concept of ecosystem services. Such information may be useful to promote or enhance dialogue among farmers, scientists and managers.
• 5. These results may contribute to the devising of ecosystem management measures in the Negro River, aimed to conserve both rivers and their associated floodplain forests, involving local fishermen. In the Atlantic Forest, we proposed some initiatives, such as to allow direct economic use of their forests to conciliate conflicting perceptions of farmers about ecological benefits versus economic losses from reforestation. Despite their cultural, environmental and geographical differences, the two study cases are complementary and cost-effective and promising approaches to including LEK in the design of ecological research.

Copyright © 2007 John Wiley & Sons, Ltd.