Distribution of muscarinic receptor subtypes and interstitial cells of Cajal in the gastrointestinal tract of healthy dairy cows

OBJECTIVE: To describe the distribution of muscarinic receptor subtypes M(1) to M(5) and interstitial cells of Cajal (ICCs) in the gastrointestinal tract of healthy dairy cows. SAMPLE POPULATION: Full-thickness samples were collected from the fundus, corpus, and pyloric part of the abomasum and from the duodenum, ileum, cecum, proximal loop of the ascending colon, and both external loops of the spiral colon of 5 healthy dairy cows after slaughter. PROCEDURES: Samples were fixed in paraformaldehyde and embedded in paraffin. Muscarinic receptor subtypes and ICCs were identified by immunohistochemical analysis. RESULTS: Staining for M(1) receptors was found in the submucosal plexus and myenteric plexus. Antibodies against M(2) receptors stained nuclei of smooth muscle cells only. Evidence of M(3) receptors was found in the lamina propria, in intramuscular neuronal terminals, on intermuscular nerve fibers, and on myocytes of microvessels. There was no staining for M(4) receptors. Staining for M(5) receptors was evident in the myocytes of microvessels and in smooth muscle cells. The ICCs were detected in the myenteric plexus and within smooth muscle layers. Distribution among locations of the bovine gastrointestinal tract did not differ for muscarinic receptor subtypes or ICCs. CONCLUSIONS AND CLINICAL RELEVANCE: The broad distribution of M(1), M(3), M(5), and ICCs in the bovine gastrointestinal tract indicated that these components are likely to play an important role in the regulation of gastrointestinal tract motility in healthy dairy cows. Muscarinic receptors and ICCs may be implicated in the pathogenesis of motility disorders, such as abomasal displacement and cecal dilatation-dislocation.     

Association between serum cobalamin and methylmalonic acid concentrations in dogs

The aim of this study was to evaluate the association between serum methylmalonic acid (MMA), a proposed marker of cellular cobalamin deficiency, and serum cobalamin concentrations in dogs. Serum samples from 555 dogs were grouped according to their serum cobalamin concentrations (<150 ng/L to 1000 ng/L). Additionally, serum samples were collected from 43 healthy dogs to calculate a reference interval for canine serum MMA. MMA was measured using a GC/MS method. Groups were compared using a Kruskal-Wallis test with Dunn's post test. Proportions of dogs above the upper limit of the reference interval were calculated and a χ2-test for trend was performed to evaluate the association between serum cobalamin and MMA concentrations. The reference interval for serum MMA was calculated to be 414.7-1192.5 nmol/L. Dogs with serum cobalamin concentrations <251 ng/L had significantly higher MMA concentrations (P<0.05) and the χ2-test for trend showed a trend for increasing serum MMA concentrations with decreasing serum cobalamin concentrations (P<0.0001). Additionally, a number of dogs with normal serum cobalamin concentrations had increased serum MMA concentrations, suggesting that some of these dogs may have cobalamin deficiency on a cellular level. Further studies are warranted to determine if these dogs should receive cobalamin supplementation.     

Genotyping of Bacillus anthracis Isolated from Croatia and Bosnia and Herzegovina

Anthrax is a serious disease caused by Bacillus anthracis. Humans can become infected by handling products from infected animals, by breathing spores and rarely by eating undercooked meat from infected animals. The genome of B. anthracis is highly monomorphic and thus shows very low DNA sequence variation. We analysed the molecular characteristics of 12 B. anthracis isolates from outbreaks in Croatia and Bosnia and Herzegovina, which have occurred during the past 10 years along with two vaccine strains. Genotyping system based on variable‐number tandem repeat analysis at six loci revealed that six isolates belong to genotype from the A1.a cluster whilst six isolates relate to the B2 cluster, compared to 89 previously described genotypes. The distribution of two evolutionarily distant clusters suggests an introduction of B. anthracis to this area in at least two separate events.     

Nitrogen retention and plant uptake on a highly weathered central Amazonian Ferralsol amended with compost and charcoal

Leaching losses of N are a major limitation of crop production on permeable soils and under heavy rainfalls as in the humid tropics. We established a field trial in the central Amazon (near Manaus, Brazil) in order to study the influence of charcoal and compost on the retention of N. Fifteen months after organic‐matter admixing (0–0.1 m soil depth), we added ¹⁵N‐labeled (NH₄)₂SO₄ (27.5 kg N ha^–1 at 10 atom% excess). The tracer was measured in top soil (0–0.1 m) and plant samples taken at two successive sorghum (Sorghum bicolor L. Moench) harvests. The N recovery in biomass was significantly higher when the soil contained compost (14.7% of applied N) in comparison to only mineral‐fertilized plots (5.7%) due to significantly higher crop production during the first growth period. After the second harvest, the retention in soil was significantly higher in the charcoal‐amended plots (15.6%) in comparison to only mineral‐fertilized plots (9.7%) due to higher retention in soil. The total N recovery in soil, crop residues, and grains was significantly (p < 0.05) higher on compost (16.5%), charcoal (18.1%), and charcoal‐plus‐compost treatments (17.4%) in comparison to only mineral‐fertilized plots (10.9%). Organic amendments increased the retention of applied fertilizer N. One process in this retention was found to be the recycling of N taken up by the crop. The relevance of immobilization, reduced N leaching, and gaseous losses as well as other potential processes for increasing N retention should be unraveled in future studies.     

Charcoal and smoke extract stimulate the soil microbial community in a highly weathered xanthic Ferralsol

The influence of charcoal and smoke condensates (pyroligneous acid, PA) on microbial activity in a highly weathered Amazonian upland soil was assessed via measurements of basal respiration (BR), substrate-induced respiration (SIR), and exponential population increase after substrate addition. PA extracts are commonly used for fertilizer or as pest control in Brazil, where phosphorus (P) availability and nitrogen (N) leaching are among the most severe limitations for agriculture. Microbes play an important role in nutrient cycling and solubilizing of phosphate. BR, microbial biomass, population growth and the microbe's efficiency (expressed by the metabolic quotient) increased linearly and significantly with increasing charcoal concentrations (50, 100 and 150 g kg⁻¹ soil). Application of PA caused a sharp increase in all parameters. We suppose that the condensates from smoke contain easily degradable substances and only small amounts of inhibitory agents, which could be utilized by the microbes for their metabolism.     

Prevalence and identification of fungal DNA in the small intestine of healthy dogs and dogs with chronic enteropathies

Limited information is available about the prevalence and phylogenetic classification of fungal organisms in the gastrointestinal tract of dogs. Also, the impact of fungal organisms on gastrointestinal health and disease is not well understood. The aim of this study was to evaluate the prevalence of fungal DNA in the small intestine of healthy dogs and dogs with chronic enteropathies. Small intestinal content was analyzed from 64 healthy and 71 diseased dogs from five different geographic locations in Europe and the USA. Fungal DNA was amplified with panfungal primers targeting the internal transcriber spacer (ITS) region. PCR amplicons were subjected to phylogenetic analysis. Fungal DNA was detected in 60.9% of healthy dogs and in 76.1% of dogs with chronic enteropathies. This prevalence was not significantly different between the two groups (p=0.065). Fungal DNA was significantly more prevalent in mucosal brush samples (82.8%) than in luminal samples (42.9%; p=0.002). Sequencing results revealed a total of 51 different phylotypes. All sequences belonged to two phyla and were classified as either Ascomycota (32 phylotypes) or Basidiomycota (19 phylotypes). Three major classes were identified: Saccharomycetes, Dothideomycetes, and Hymenomycetes. The most commonly observed sequences were classified as Pichia spp., Cryptococcus spp., Candida spp., and Trichosporon spp. Species believed to be clinically more important were more commonly observed in diseased dogs. These results indicate a high prevalence and diversity of fungal DNA in the small intestine of both healthy dogs and dogs with chronic enteropathies. The canine gastrointestinal tract of diseased dogs may harbor opportunistic fungal pathogens.