Investigation of Interspecies Interactions within Marine Micromonosporaceae Using an Improved Co-Culture Approach

With respect to bacterial natural products, a significant outcome of the genomic era was that the biosynthetic potential in many microorganisms surpassed the number of compounds isolated under standard laboratory growth conditions, particularly among certain members in the phylum Actinobacteria. Our group, as well as others, investigated interspecies interactions, via co-culture, as a technique to coax bacteria to produce novel natural products. While co-culture provides new opportunities, challenges exist and questions surrounding these methods remain unanswered. In marine bacteria, for example, how prevalent are interspecies interactions and how commonly do interactions result in novel natural products? In an attempt to begin to answer basic questions surrounding co-culture of marine microorganisms, we have tested both antibiotic activity-based and LC/MS-based methods to evaluate Micromonosporaceae secondary metabolite production in co-culture. Overall, our investigation of 65 Micromonosporaceae led to the identification of 12 Micromonosporaceae across three genera that produced unique metabolites in co-culture. Our results suggest that interspecies interactions were prevalent between marine Micromonosporaceae and marine mycolic acid-containing bacteria. Furthermore, our approach highlights a sensitive and rapid method for investigating interspecies interactions in search of novel antibiotics, secondary metabolites, and genes.     

Quantification of reservoir bags as airway pressure-limiting devices in a nonrebreathing system

ObjectiveTo evaluate the influence of reservoir bag types, volumes and previous use on the peak pressures (Pmax) and the times to develop 30 cmH₂O pressure (P30) within a nonrebreathing system with a closed adjustable pressure-limiting (APL) valve.Study designIn vitro study using three-way factorial design with repeated measure on one factor.SubjectsA total of 75 new anesthesia reservoir bags (five types, three volumes, five bags from each type × volume). The bag types were reusable latex (RL), disposable latex (DL) and three disposable neoprene (DN-1, DN-2 and DN-3).MethodsEach bag was tested three times (treatments): new, after prestretching and 1 week later. The bags were attached to a Bain system and anesthesia machine with closed APL valve and patient port with O₂ flow 2 L minute^–1 until Pmax was reached. The Pmax and time to reach P30 values were determined from recorded pressure traces. General linear mixed model analysis was used to examine the effects of bag type, volume and treatment. One-sided 95% upper prediction limits of Pmax were calculated to test the null hypothesis that predicted Pmax of new bags would be ≥ 50 cmH₂O for each factor combination.ResultsRL bags were the least and DN-3 bags were the most compliant. Prestretching increased compliance in all bag types. Smaller bags of RL, DL and DN-1 were less compliant than larger ones. The predicted Pmax values were < 50 cmH₂O only for DN-3 bags after prestretching. The time to reach P30 was critically low when using 0.5 L bags (median 17 seconds).Conclusions and clinical relevanceTo minimize the risk of barotrauma, highly compliant reservoir bags (e.g. DN-3) are recommended and reusable bags should be avoided. Bags should be prestretched before first use, 0.5 L bags should be avoided and fresh gas flow minimized.     

Correlation between cytopathology and histopathology of the skin, lymph node and spleen in 500 dogs and cats

The results of 382 cytological examinations in dogs and 118 in cats were retrospectively compared with the correlating histological results.The histological diagnosis represented the gold standard. The investigation comprised skin, lymph node and spleen samples. The aim was to estimate the diagnostic value and the limits of cytological examinations in relation to the type of the changes. The cases were grouped in six categories: Corresponding diagnoses were formulated in 201 cases. In lymph node samples, they were reached almost twice as frequently than in skin and spleen samples. Round cell neoplasias (mast cell tumours, canine cutaneous histiocytomas and malignant lymphomas) were most often subclassified. Cytological diagnoses lacking subclassification were reached in 98 cases. The lesions were cytologically less precisely characterized than histologically. Provisional cytological diagnosis or differential diagnosis were formulated in 112 cases. Most commonly, the distinction between neoplastic and non-neoplastic character of a lesion was not possible (especially in mesenchymal spindle-cell proliferations). In 53 cases, the cytological diagnosis could not be established. In 22 cases, the relevant lesion was diagnosed only histologically whereas the cytological picture revealed another, usually secondary tissue reaction. Cytologically incorrect diagnoses were formulated in 14 cases.     

Dislodgeable residues and persistence of diazinon,chlorpyrifos and isofenphos following their application to turfgrass

In laboratory experiments, residues of diazinon applied to turfgrass, Poa pratensis L., that could be dislodged by rubbing with cheese-cloth, declined from c. 10% of the total applied when the grass was vigorously rubbed immediately after application to 0.3% after 1 day. Sunlight did not influence the rate of decline in dislodgeable residues or residues remaining on or within the leaf blades. In field experiments where 4.5 kg ha^?1 of diazinon was applied in liquid or granular form, about 20 times more diazinon was dislodged from the liquid formulation immediately after application than from the granular. By 1 day after application the percentage of the total applied diazinon that could be dislodged was equal for both formulations. Rainfall had a significant effect on the amount dislodged from grass blades, but mowing did not. Similar rates of decline in the dislodgeable fraction of diazinon, chlorpyrifos and isofenphos were observed in field experiments. Recovery of the dislodgeable fraction declined to 0.25% or less of the total amount of any of these insecticides by 1 day after application. However, residues in the thatch remained sufficiently high for control of insects for up to 7 days after application for diazinon and 14 days for chlorpyrifos and isofenphos.     

Identification of focal adenomyosis as a uterine lesion in two dogs

A focal uterine adenomyosis is described in two bitches. In both cases, the uterus showed knobbly enlargements of 4 to 8 cm in diameter, which resulted in distinct clinical symptoms. Other pathological changes of the uterus were not present. One bitch was presented because of a history of vaginal discharge of several months' duration. Radiographs, as well as ultrasonography, revealed a soft tissue lesion at the cervix. The other bitch showed a marked reduction in its general condition and a sudden onset of a tense abdomen. Radiologically, a lesion of soft tissue opacity was observed in the mid-abdomen and was seen to originate from the left uterine horn during exploratory laparotomy. A torsion of the lesion was present, which explained the clinical signs in this second case.     

The influence of a fermentation-resistant glucose diet on the glucose concentration and other metabolites in portal and jugular blood in cows

The goal of this study was to determine the effect of fermentation-resistant glucose on the glucose concentration and other metabolites in portal and jugular blood in 15 non-lactating cows. In all cows, an indwelling catheter was placed in the left jugular vein and the portal vein for collection of blood samples. Five control cows were fed hay as a normal diet, five control cows were fed straw to induce an energy deficit and five cows were fed hay and they received additionally 2000 g of a fermentation-resistant D-glucose product. The glucose concentration in jugular and portal blood was not influenced by feeding. The concentration of urea and bile acids were significantly higher in portal blood than jugular blood. There was no difference between portal and jugular blood of glucose and total solids. Diet had a significant effect on the concentrations of ammonia, urea, free fatty acids and triglycerides. The concentrations of ammonia and urea were higher in blood of cows fed straw than in blood of cows fed either hay or a fermentation-resistant glucose product. The concentration of urea remained constant in cows fed hay, but increased in cows fed straw and decreased in cows fed a fermentation-resistant glucose product. The concentration of free fatty acids and triglycerides were significantly higher in cows fed a fermentation-resistant glucose product than in cows fed hay. In the present study, a single administration of 300 g of fermentation-resistant glucose did not affect the concentration of blood glucose. Therefore, despite ongoing promotion of such products, there is no indication at this time that administration of fermentation-resistant glucose to cows at the start of lactation results in an increase in blood glucose concentration.     

Ultrasonography in gastrointestinal disease in cattle

Ultrasonography is an ideal diagnostic tool for investigating gastrointestinal disorders in cattle. It is performed on standing non-sedated cattle using a 3.5 MHz linear transducer. In animals with traumatic reticuloperitonitis, inflammatory fibrinous changes, and abscesses can be imaged; however, magnets and foreign bodies are difficult to visualize because of the gas content of the reticulum. Ultrasonography can be used to assess the size, position and contents of the abomasum. Percutaneous ultrasound-guided abomasocentesis can be performed to evaluate the nature and chemical composition of its contents. In left displacement of the abomasum, the abomasum is seen between the left abdominal wall and the rumen. It contains fluid ingesta ventrally and a gas cap of varying size dorsally. Occasionally, the abomasal folds are seen in the ingesta. In cattle with right displacement of the abomasum, the liver is displaced medially from the right abdominal wall by the abomasum, which has an ultrasonographic appearance similar to that described for left displacement. Motility and diameter of the intestine are the most important criteria for ultrasonographic assessment of ileus. However, the cause of the ileus is rarely determined using ultrasonography. In cases with ileus of the small intestine, there is at least one region of dilatation of the intestine and motility is reduced or absent. In cattle with caecal dilatation, the caecum can always be imaged from the right lateral abdominal wall. The wall of the caecum closest to the transducer appears as a thick, echogenic, semi-circular line.     

Assessment of a pyrogallol red technique for total protein measurement in the cerebrospinal fluid of dogs

The measurement of protein concentration in the cerebrospinal fluid is a basic analytical method in neurology. In this study, a pyrogallol red technique using a human albumin calibrator previously validated in human medicine was tested for canine samples, and the results were compared with those obtained using urine test strips. Pyrogallol red significantly (P<0.05) but moderately underestimated purified dog albumin and globulins. The imprecision of the technique was low: intra- and between-series coefficients of variation were 1.6 and 4.3 per cent at protein concentrations of about 0.3 g/litre. Over 49 samples, there was good agreement between the pyrogallol red and test strip results (r=0.63), especially for low and high protein concentrations, but misclassifications were observed with '+' test strip readings.