Percutaneous absorption of chemicals: developing an understanding for the treatment of disease in frogs

The permeable nature of frog skin presents an alternative route for the delivery of therapeutic chemicals to treat disease in frogs. However, although therapeutic chemicals are often topically applied to the skin of frogs, their pharmacokinetics have rarely been reported. To provide evidence to guide both candidate drug and formulation selection, we highlight factors expected to influence percutaneous absorption through frog skin, including the anatomy and physiology of the skin and the physicochemical properties of applied therapeutic chemicals. Importantly, we also highlight the effects of the formulation on percutaneous absorption, especially the inclusion of potential penetration enhancers as excipients. Finally, we collate empirical data on the topical application of various therapeutic chemicals in postmetamorphic frogs and show that, in contrast to mammalian species, even large chemicals (i.e. >500 Da) and those with a wide range of log P values (?4 through +6) are likely to be absorbed percutaneously. Topical application in frogs thus promises a convenient and effective method for delivering systemic treatments of a diverse range of chemicals; however, further experimental quantification is required to ensure optimal outcomes.     

Comparison of Blood Non‐Specific Immune Parameters in Bovine Virus Diarrhoea Virus (BVDV) Persistently Infected and in Immune Heifers

Several data from different authors show that Bovine virus diarrhoea virus (BVDV) could be a key component in multiple‐etiology diseases, indeed a lower leukocytes number and their impaired functions decrease the resistance to infections. However, most of the information on the impairment of immune function during BVDV infections arise from circumstantial evidence and from experimental infection studies, and few from field data. To assess the effects of BVDV on blood cells parameters, cellular and humoral functions under field conditions, we designed a controlled study in commercial dairy herds, comparing persistent infected (PI) and healthy heifers. A total of 45 heifers were considered, the PI animals were nine, the control animals were 34, while two controls were considered as acute infected animals. The comparison of the mean values in PI calves showed a significant decrease for leukocytes and granulocytes, while platelets showed a significant increase, when compared with control animals. The total number of lymphocytes decreased not significantly in PI animals, while the proportion significantly increased. The number and proportion of monocytes was significantly reduced in PI animals, when compared with controls. The data collected on markers of cellular immunity during our study cannot be compared with the literature because there are no reference values. The presence of a persistent infection affected the cellular enzymes: NAGase, lysozyme and respiratory burst showed a large statistically significant decrease in PI animals when compared with controls. The presence of a persistent infection with BVD virus influenced blood cells number and impaired some blood cell functions. Such impairment confirms that PI animals represent a threat to the herd not only because they could spread BVDV, but also because they are more susceptible to other infectious diseases.     

Effect of floor cooling on performance of lactating sows during summer

Forty Landrace × Large White lactating sows were used to evaluate the effects of cooling of the floor when maintained under high temperature conditions during summer on their productive and reproductive performance. The sows were allocated in a completely randomized design with two treatments with 20 replicates according to parity number and body weight, with each animal being considered an experimental unit. The treatments consisted of cooling of the floor under the sow with water circulation at about 17 °C and no cooling. The resulting temperatures of the floor were 27.6 and 35.8 °C for the cooled and the control treatments, respectively. The sows from both treatments were exposed to average maximum and minimum environmental temperatures of 26.9 and 20.8 °C, respectively. Sows maintained on a cooled floor had a higher feed intake (6.47 vs. 5.61 kg/day; P < 0.01). Despite this higher intake, sows maintained on a cooled floor had higher body weight and body protein losses during the lactation period (P < 0.01) in connection with a higher milk yield and subsequent growth of the litter (2280 vs. 1798 g/day; P < 0.01). There was an effect of treatment on rectal temperature, surface temperatures and respiratory rate (P < 0.01) with lower values in sows submitted to floor cooling. It is concluded that floor cooling under the lactating sow improves its productive and reproductive performance, as well as the performance of its litter.     

The clinical, cerebrospinal fluid, and histopathologic effects of epidural ketorolac in dogs

OBJECTIVE: To evaluate the clinical, cerebrospinal fluid (CSF), and histopathologic effects of epidural ketorolac. STUDY DESIGN: Blinded, randomized, placebo controlled study. ANIMALS: Twenty-two adult mixed breed dogs with 16 treatment and 6 control dogs, weighing 14.4 to 29.8 kg. METHODS: Dogs were anesthetized and epidural catheters were placed at the lumbosacral space. Catheter placement was evaluated fluoroscopically. Ketorolac (0.4 mg/kg) or placebo (5% ethanol) was administered epidurally over a 52-hour period, with 5 injections given at 12-hour intervals. At 1, 2, 4, or 8 hours after the first and last injection of ketorolac, dogs were anesthetized and CSF was obtained. Control dogs had CSF sampled 1 hour after the first and last ethanol injection. Neurologic function and pain responses were evaluated before and during the study. Selected dogs were then killed and necropsies performed. RESULTS: None of the dogs exhibited any clinical or neurologic abnormalities during the study. No statistical difference was noted in pain response or CSF analysis between treatment and control dogs. Gross necropsy revealed gastrointestinal ulceration of varying degrees in all treatment dogs. Histopathologic analysis of the spinal cord and meninges revealed minimal focal leptomeningeal phlebitis in 2 of 8 treatment dogs and minor subdural inflammation in 1 control dog. No changes to the neural structures were noted in any dogs. CONCLUSIONS: Epidural administration of ketorolac did not cause clinical signs, alteration in CSF values, or pathologic changes to the spinal cord when used for short duration. Gastrointestinal ulceration was common when ketorolac was administered epidurally at 0.4 mg/kg every 12 hours for 5 treatments. CLINICAL RELEVANCE: This study documented the neurologic safety of epidural ketorolac in dogs before an efficacy trial can be performed. Gastrointestinal ulceration may limit use to short duration or a single injection.     

Incomplete systemic movement of Xanthomonas campestris pv. musacearum and the occurrence of latent infections in xanthomonas wilt‐infected banana mats

Management of banana xanthomonas wilt (XW) (caused by Xanthomonas campestris pv. musacearum, Xcm) has been impeded by poor adoption of control options that are complex, cumbersome and costly. To improve XW management, this study investigated Xcm survival and latent infections in subsequent generations, survival of latently infected planting materials (suckers), incidence of latent infections in symptomless plants in mats having diseased plants, and XW status across farms and markets in districts previously devastated but currently endemic. On‐station experiments were protected from new infections. Latent bacteria at low levels were detected in up to 20% of the third generation suckers, with a significant (P < 0·05) reduction (43–20%) in subsequent generations. Only 3–6% of latently infected suckers succumbed to XW. Incidence of Xcm in symptomless suckers from farmers' fields (with up to 70% incidence) was low (3%) while it increased (8–25%) with disease severity in mats in controlled experiments. In the surveyed districts, incidence had significantly declined with yields observed to have recovered relative to earlier reports, although latent infections remained high. This study provides evidence that if new infections are prevented, fields with high XW incidence can be rejuvenated. It showed incomplete systemic movement of Xcm in mats coupled to a gradual decline of bacterial load in subsequent generations to levels that cannot initiate disease. These studies explain the current successes in farms practising single diseased plant removal instead of whole mat rouging, and gives hope to farmers lacking access to clean planting material.     

Rumen odd and branched chain fatty acids in relation to in vitro rumen volatile fatty acid productions and dietary characteristics of incubated substrates

A first aim of this batch in vitro experiment (21 h) was to use changes in odd and branched chain fatty acid (OBCFA) patterns to suggest shifts in microbial populations, associated with four types of incubated whole dairy cow diets. Principal component analysis suggested higher dietary starch increased the proportion of C15:0 and C17:0, whereas increased neutral detergent fibre content was positively related to anteiso C15:0 concentrations, which is in agreement with the importance of these fatty acids in respectively amylolytic and cellulolytic bacteria. A second aim of the experiment was to relate rumen volatile fatty acid proportions to OBCFA by principal component regression and to compare these relations with predictions based on diet proximate composition. The R2 values achieved for the regressions between acetate, propionate and butyrate, and OBCFA were 79.6%, 86.6% and 84.9% respectively. Moreover, in the current study, predictions of the rumen fermentation pattern showed higher R2 (p < 0.01) when based on OBCFA compared with proximate feed composition. If relations persist in vivo, there could be scope for milk OBCFA to predict the supply of specific rumen nutrients.     

Exploration of plant genomes in the FLAGdb++ environment

Background

Efficient high throughput screening systems of useful mutants are prerequisite for study of plant functional genomics and lots of application fields. Advance in such screening tools, thanks to the development of analytic instruments. Direct analysis in real-time (DART)-mass spectrometry (MS) by ionization of complex materials at atmospheric pressure is a rapid, simple, high-resolution analytical technique. Here we describe a rapid, simple method for the genetic discrimination of intact Arabidopsis thaliana mutant seeds using metabolic profiling by DART-MS.

Results

To determine whether this DART-MS combined by multivariate analysis can perform genetic discrimination based on global metabolic profiling, intact Arabidopsis thaliana mutant seeds were subjected to DART-MS without any sample preparation. Partial least squares-discriminant analysis (PLS-DA) of DART-MS spectral data from intact seeds classified 14 different lines of seeds into two distinct groups: Columbia (Col-0) and Landsberg erecta (Ler) ecotype backgrounds. A hierarchical dendrogram based on partial least squares-discriminant analysis (PLS-DA) subdivided the Col-0 ecotype into two groups: mutant lines harboring defects in the phenylpropanoid biosynthetic pathway and mutants without these defects. These results indicated that metabolic profiling with DART-MS could discriminate intact Arabidopsis seeds at least ecotype level and metabolic pathway level within same ecotype.

Conclusion

The described DART-MS combined by multivariate analysis allows for rapid screening and metabolic characterization of lots of Arabidopsis mutant seeds without complex metabolic preparation steps. Moreover, potential novel metabolic markers can be detected and used to clarify the genetic relationship between Arabidopsis cultivars. Furthermore this technique can be applied to predict the novel gene function of metabolic mutants regardless of morphological phenotypes.