The organophosphorous insecticide chlorpyrifos affects the neuroepithelial junction, the bioelectric parameters of the skin of the frog Caudiverbera caudiverbera, and the structure of model cell membranes

The current study examines the acute effects of the organophosphorus pesticide chlorpyrifos on a sympathetic synapse of the frog Caudiverbera caudiverbera. Nerve stimulation was followed immediately by a transient increase in the short-circuit current (SCC) and in the potential difference (PD), which consisted of a rapid and then a slow component. Chlorpyrifos concentrations from 5 μM to 1.0 mM caused a dose-dependent block of both components to a 10% of their control values, which was reversed by washout. The pesticide blockade did not affect the skin response to noradrenaline. X-ray diffraction and fluorescence spectroscopy studies on membrane models showed marked phospholipid perturbation, which favors changes in ion channel conformation and interferes with receptor proteins, thus altering noradrenergic transmission and Cl⁻ secretion in the mucous glands. The foregoing results may be interpreted as a reversible inhibition of the neuroepithelial synapse to nerve stimulation, possibly due to non-specific lip μd-protein perturbation, interference with synaptic transmission, and transient Cl⁻ channel inactivation.     

A potential early clinical phenotype of necrotizing meningoencephalitis in genetically at‐risk pug dogs

BackgroundNecrotizing meningoencephalitis (NME) in the pug dogs is a fatal neuroinflammatory disease associated with rapid progression and poor response to conventional immunosuppressive therapy. Diagnosis is typically made after severe neurological abnormalities have manifested.Hypothesis/ObjectivePug dogs at genetic risk for NME might manifest neurological abnormalities before developing pathognomonic clinical signs of NME.AnimalsThirty‐six pug dogs less than 4 years of age asymptomatic for NME.MethodsProspective observational cohort study with germline genome‐wide genotyping. Neurological examinations were performed 4 weeks apart to document reproducible findings of central nervous system disease. Magnetic resonance imaging, cerebrospinal fluid analysis, and testing for infectious diseases were performed in all pugs with reproducible abnormalities detected on neurological examination.ResultsThe overall risk allele frequency in this cohort was 40%; 5 (14%) dogs were high risk, 19 (53%) dogs were medium risk, and 12 (33%) dogs were low genetic risk for NME. Reproducible abnormalities detected on neurological examination were identified in 8/24 (33%) genetically at‐risk dogs and 0/12 (0%) low risk dogs. Clinical abnormalities included multifocal spinal pain in 8/8, reduced menace response in 5/8, and lateralizing postural reaction deficits in 5/8 pugs. There was a strong association between genotype risk and the presence of this clinical phenotype (P = .03).Conclusions and Clinical ImportanceOur findings suggest the presence of a novel early clinical phenotype of NME in apparently asymptomatic genetically at‐risk pugs which might be used to plan early diagnostic and therapeutic clinical trials.     

Recruitment of catarina scallop (Argopecten ventricosus) larvae on artificial collectors off the NE coast of the Gulf of California

We evaluated recruitment of larvae of catarina scallop, Argopecten ventricosus, in the area of Puerto Peñasco, NE Gulf of California. We moored artificial collectors in six sites from June 2007 to August 2008 and replaced them every 2 months. We used monthly (July 2002–September 2011) sea surface temperature (SST, °C) and surface chlorophyll‐a concentration (SSChl, mg m^?3) Aqua/MODIS satellite data to describe seasonal environmental behaviour study area. Also, we recorded bottom temperature at each site every 4 h, and every 2 months measured sea surface salinity, temperature and dissolved oxygen. We used a repeated measures anova to evaluate differences in the number of recruited spat between main factors, and analysed the presence of multimodal spat shell size frequency distributions. Overall, spat recruitment was negatively correlated with seawater temperature and showed higher spat recruitment abundances throughout winter, which is the season with the highest surface chlorophyll a concentration. We estimated multimodal shell size frequency distributions characterized by more than one modal size. The natural collection of A. ventricosus spat on artificial collectors in the area can be successfully performed over a protracted period (November–December to May–June). Our results extend the area where collection of A. ventricosus spat can be successful.     

Genetic analysis of Aequipecten opercularis and Mimachlamys varia (Bivalvia: Pectinidae) in several Atlantic and Mediterranean localities, revealed by mitochondrial PCR-RFLPs: a preliminary study

Aequipecten opercularis (Queen scallop) and Mimachlamys (Chlamys) varia (Black scallop) are important natural resources occurring in Atlantic and Mediterranean coasts. To develop an optimal sustainable exploitation plan, it is important to study the genetic structure of the different populations. In this study, we used polymerase chain reaction‐restriction fragment length polymorphisms for the determination of the genetic variation and population structure of these two species in different localities. Ten composite haplotypes were generated for A. opercularis and 15 haplotypes for M. varia. Of these, six and four were unique respectively. The analysis of the distribution of the different haplotypes between the localities showed no clear evidence of subdivision in A. opercularis, while in M. varia the results indicated that the two localities analysed should be managed as separate stocks.     

Residue level, persistence, and storage performance of citrus fruit treated with fludioxonil

The potential of postharvest dip treatments with fludioxonil (FLU) (a synthetic analogue of the bacterial metabolite of pyrrolnitrin), in controlling postharvest decay caused by Penicillium digitatum and Penicillium italicum of citrus fruit was investigated in comparison with the conventional fungicide imazalil (IMZ). The ultrastructural changes of fruit epicuticular wax was investigated as a function of water dip temperature, and the possible role of these changes was related to residue accumulation under FLU treatment. Residues retained by fruit were determined as a function of fungicide concentration, dip temperature, and fruit storage conditions. Scanning electron microscopy analysis revealed that fruit dipping in water at 30 or 40 degrees C did not cause differences in cuticular wax's ultrastructure in comparison to control fruit, while treatments at 50, 55, or 60 degrees C caused the disappearance of wax platelets, resulting in relatively homogeneous skin surface, due to partial "melting" of epicuticular wax. Residues of FLU in fruit treated at 20 or 50 degrees C were significantly correlated with the doses of fungicide applied. When equal amounts of fungicide were employed, the residue concentrations were notably higher (from 2.6- to 4-fold) in fruit treated at 50 degrees C than in fruit treated at 20 degrees C. The dissipation rate of FLU in "Salustiana" and "Tarocco" oranges was lower in fruit subjected to treatment at 50 degrees C. The minimal FLU concentration for almost complete decay control in artificially wounded fruit during 7-d storage at 20 degrees C was 400 mg/L active ingredient (ai) in fruit treated at 20 degrees C and 100 mg/L ai in fruit treated at 50 degrees C. Results on nonwounded Tarocco oranges subjected to 3 weeks of simulated quarantine conditions at 1 degrees C, plus 6 weeks of standard storage at 8 degrees C and an additional two weeks of simulated marketing period (SMP) at 20 degrees C revealed that almost complete decay control with FLU applications of 100 mg/L at 50 degrees C and 400 mg/L at 20 degrees C resulted in ca. 0.8 mg/kg FLU fruit residues, in agreement with results on wounded citrus fruit. When equal concentrations and temperatures were applied, FLU treatments were as effective as IMZ. In vitro trials showed a low sensitivity to FLU against P. digitatum and P. italicum isolates. MIC values for the complete inhibition of mycelium growth were >or=100 microg/mL, while ED(50) values ranged from 0.1 to 1 microg/mL for P. digitatum and from 1 to >100 microg/mL for P. italicum. The latter result suggests that care should be taken to avoid exclusive application of FLU in a sustainable program for management of fruit decay. However, integrating fungicide application and hot water dip may reduce the possibility of selecting fungicide-resistant populations of the pathogen, by increasing the effectiveness of the treatment.     

Disturbance to a foraging seabird by sea-based tourism: Implications for reserve management in marine protected areas

The provision of recreational opportunities is one of the important human goals of marine protected areas. However, as levels of recreational use increase, human disturbance is likely to cause significant detrimental effects upon wildlife. Here we evaluate the best managing options to mitigate the impact of sea-based tourism on the foraging activity of an endangered population of European shags, Phalacrocorax aristotelis, in a coastal marine protected area (Cíes islands, north-western Iberia). Boat disturbance elicited a characteristic avoidance behavior that resulted in a substantial reduction in foraging activity as levels of boat use increased. Moreover, boats excluded shags from the best feeding areas, resulting in higher densities of foragers in areas of little boat traffic. We used a behavioral model to explore the effects of managing strategies aimed at reducing the impact of boats on the foraging activity of shags. Our model suggested that in low boat disturbance scenarios limiting the number of boats using the reserve would be a better management option than habitat protection (i.e. the establishment of set-aside areas free of boat traffic). On the contrary, when boat disturbance levels are high the protection of habitat is recommendable, even if spatial variation in habitat quality is unknown or poorly assessed. Our study stresses the point that management strategies to minimize disturbance to foraging seabirds may depend on the spatial overlap between sea-based recreational activities and foraging seabirds and the spatial variation in marine habitat quality for seabirds.     

Genetic immunization with the immunodominant antigen P48 of Mycoplasma agalactiae stimulates a mixed adaptive immune response in BALBc mice

A DNA vaccine against contagious agalactia was developed for the first time, encoding the P48 of Mycoplasma agalactiae. Specific immune responses elicited in BALB/c mice were evaluated. Both total IgG and IgG1 were detected in mice vaccinated with pVAX1/P48. Proliferation of mononuclear cells of the spleen, levels of gamma interferon, interleukin-12, and interleukin-2 mRNAs were enhanced in immunized animals. Results indicate that pVAX1/P48 vaccination induced both T_h1 and T_h2 immune responses. Nucleic acid immunization could be a new strategy against M. agalactiae infections and may be potentially used to develop vaccines for other Mycoplasma diseases.     

Salmonella enterica serovar Choleraesuis derivatives harbouring deletions in rpoS and phoP regulatory genes are attenuated in pigs, and survive and multiply in porcine intestinal macrophages and fibroblasts, respectively

Live attenuated Salmonella enterica strains have been extensively studied as potential vectors for the oral delivery of heterologous antigens. Due to its ability to target immune cells, its specific mechanism for crossing the intestinal barrier, and its swine-restricted tropism, S. enterica subspecies enterica serovar Choleraesuis (S. Choleraesuis) has attracted a great deal of interest for the production of bacterial-based oral carriers specifically adapted to swine. In this study, two mutants of S. Choleraesuis were constructed and their attenuation and intracellular fate analysed with the purpose of engineering new attenuated live strains with improved properties as oral vaccine carriers. Those strains harboured a specific deletion either within the phoP or rpoS genes, which encode virulence-related regulators in S. Typhimurium. In comparison to the wild-type parental S. Choleraesuis, the mutant strains, especially DeltaphoP, were extremely low in virulence in the murine model and in the natural host, the pig. Moreover, when compared with a commercial live vaccine strain, SC-54, the two mutants showed a higher level of attenuation in mice and DeltaphoP also in pigs. In addition, DeltarpoS and DeltaphoP presented a proliferation and survival phenotype within swine intestinal primary fibroblast and macrophage cell cultures, respectively. Collectively, the present results indicate that the DeltarpoS and DeltaphoP strains of S. Choleraesuis gather adequate features to be potential candidates for vaccine vectors for the specific delivery of heterologous antigens adapted to pigs.     

Pharmacokinetics of a combination preparation of ampicillin and sulbactam in turkeys

OBJECTIVE: To investigate the disposition kinetics of ampicillin and sulbactam after IV and IM administration of an ampicillin-sulbactam (2:1) preparation and determine the bioavailability of the combined preparation after IM administration in turkeys. ANIMALS: 10 healthy large white turkeys. PROCEDURE: In a crossover study, turkeys were administered the combined preparation IV (20 mg/kg) and IM (30 mg/kg). Blood samples were collected before and at intervals after drug administrations. Plasma ampicillin and sulbactam concentrations were measured by use of high-performance liquid chromatography; plasma concentration-time curves were analyzed via compartmental pharmacokinetics and noncompartmental methods. RESULTS: The drugs were distributed according to an open 2-compartment model after IV administration and a 1-compartment model (first-order absorption) after IM administration. For ampicillin and sulbactam, the apparent volumes of distribution were 0.75+/-0.11 L/kg and 0.74+/-0.10 L/kg, respectively, and the total body clearances were 0.67+/-0.07 L x kg(-1) x h(-1) and 0.56+/-0.06 L x kg(-1) x h(-), respectively. The elimination half-lives of ampicillin after IV and IM administration were 0.78+/-0.12 hours and 0.89+/-0.17 hours, respectively, whereas the corresponding half-lives of sulbactam were 0.91+/-0.12 hours and 0.99+/-0.16 hours, respectively. Bioavailability after IM injection was 58.87+/-765% for ampicillin and 53.75+/-5.35% for sulbactam. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that a regimen of loading and maintenance doses of 300 mg of the ampicillin-sulbactam (2:1) combination/kg every 8 hours could be clinically useful in turkeys. This dosage regimen maintained plasma concentrations of ampicillin > 0.45 microg/mL in turkeys.     

Identification of the core rumen bacterial taxa and their population dynamics during the fattening period in Japanese Black cattle

The rumen microbiota comprises a vast range of bacterial taxa, which may affect the production of high-quality meat in Japanese Black cattle. The aim of this study was to identify core rumen microbiota in rumen fluid samples collected from 74 Japanese Black cattle raised under different dietary conditions using 16S rRNA gene amplicon sequencing. In the rumen of fattening Japanese Black cattle, 10 bacterial taxa, showing >1% average relative abundance and >95% prevalence, irrespective of the dietary conditions and the fattening periods, were identified as the core rumen bacterial taxa, which accounted for approximately 80% of the rumen microbiota in Japanese Black cattle. Additionally, population dynamics of the core rumen bacterial taxa revealed two distinct patterns: Prevotella spp. and unclassified Bacteroidales decreased in the mid-fattening period, whereas unclassified Clostridiales, unclassified Ruminococcaceae, Ruminococcus spp., and unclassified Christensenellaceae increased during the same period. Therefore, the present study reports the wide distribution of the core rumen bacterial taxa in Japanese Black cattle, and the complementary nature of the population dynamics of these core taxa, which may ensure stable rumen fermentation during the fattening period.