Micro-environmental conditions modulate protein secretion and infectivity of the Trichinella spiralis L1 larva

After digestion of infected meat the free L1 of Trichinella spp. penetrate the intestinal mucosa where they moult to the mature adult stage. We have used proteomics to identify changes in protein secretion during in vitro culture of free T. spiralis muscle larvae under different environmental conditions, and to correlate these changes with their infectivity in mice. Muscle larvae were cultured in different media (RPMI-1640, C-199 and HBSS) under conditions of anaerobiosis, microaerobiosis and in 5% CO₂ at 37 °C. Following incubation the larval excretory/secretory proteins were analysed by two-dimensional gel electrophoresis and the larvae were used to orally infect naïve CD1 mice. For all culture media tested, infectivity of the L1 was preserved following incubation in anaerobic conditions. In contrast, the infectivity of worms cultured in nutrient-rich media was almost completely abolished in both microaerobiosis and in the presence of 5% CO₂. Some infectivity was retained in poor or reduced culture media. Comparative analysis of larval infectivity and protein secretion showed that loss of infectivity correlated with the appearance of non-tyvelosylated proteins that in turn may be related to the onset of moulting.     

Inductive expression of the NOD1 signalling pathway in chickens infected with Salmonella pullorum

1. The aim of this study was to describe the role of Nucleotide-binding oligomerization domain-containing protein 1 (NOD1) receptor signalling in chicken.

2. Tissue-specific expression analysis of NOD1, receptor-interacting serine-threonine kinase 2 (RIPK2), nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase 11 (MAPK11 or p38) by quantitative real-time PCR (qRT-PCR) revealed their wide distribution in various organs and tissues.

3. Salmonella pullorum infection activated NOD1 receptor signalling in vivo and in vitro, resulting in significant induction of downstream signalling molecules RIPK2, NF-κB/p65, MAPK11/p38 and the effector molecules IL-1b and IL-8.

4. Activation of NOD1 by its agonist bacterial γ-D-glutamyl-meso-diaminopimelic acid (iE-DAP) in HD11 cells induced the adapter molecular RIPK2 and activated the NF-κB/p65 and MAPK11/p38 pathways, resulting in an increase in IL-8 but not IL-1β. Additionally, inhibition of NOD1 using NOD1-shRNA resulted in downregulation of RIPK2, MAPK11 and IL-8, while NF-κB/p65 and IL-1β were unaltered.

5. These results highlight the important role of NOD1 receptors in eliciting the innate immune response following pathogenic invasion in chicken.     

The muscular dystrophic chicken is hypernatremic

1. The E3 ubiquitin protein ligase 1 (WWP1) gene, the mutation of which causes muscular dystrophy in chickens, is expressed not only in the pectoral muscle, but also in a number of tissues such as the kidney. Therefore, this study examined some parameters related to kidney function in muscular dystrophic (MD) chickens.

2. Plasma osmolality, Na⁺ and K⁺ concentrations, aldosterone levels, and the expression of aquaporin (AQP) 2, AQP3, and α subunits of the amiloride-sensitive epithelial sodium channel (αENaC) were analysed in the kidneys of 5-week-old MD chickens and White Leghorn (WL) chickens under physiological conditions or after one day of water deprivation.

3. Plasma osmolality, Na⁺ concentrations, and plasma aldosterone levels were significantly higher in MD chickens than in WL chickens. αENaC mRNA expression levels were lower in MD chickens than in WL chickens. AQP2 and AQP3 mRNA expression levels were similar in the two strains of chickens.

4. Plasma osmolality correlated with aldosterone levels and AQP2 and αENaC mRNA levels in WL chickens. In MD chickens, plasma osmolality correlated with AQP2 mRNA levels, but not with plasma aldosterone or αENaC mRNA levels.

5. These results suggest that neither water reabsorption nor the expression of AQP2 and AQP3 is impaired in MD chickens and that a WWP1 gene mutation may or may not directly induce an abnormality in Na⁺-reabsorption in the kidneys of MD chickens, potentially through αENaC.     

Pharmacokinetics of an injectable long‐acting parenteral formulation of doxycycline hyclate in pigs

Based on its ideal PK/PD ratios, doxycycline hyclate (DOX‐h), a time‐dependant antibacterial, is ideally expected to achieve sustained plasma drug concentrations at or slightly above the MIC level for as long as possible between dosing intervals. Pursuing this end, a poloxamer‐based matrix was used to produce a 10% long‐acting injectable preparation (DOX‐h‐LA) and its serum concentrations vs. time profile investigated after its injection to pigs in the pericaudal s.c. by parallel design. Results were compared with the forced oral bolus dose and i.v. pharmacokinetics of DOX‐h. For this study, 12 recently weaned pigs per group were included in this trial, and a dose of 20 mg/kg was injected in all cases. DOX‐h‐LA showed the greatest values for bioavailability (115.38%); maximum serum concentration (Cmax) value was 1.5 ± 0.2 with a time to reach Cmax of 3.41 ± 0.04 h and an elimination rate constant of 70.93 ± 0.87 h. Considering minimum effective serum concentration of 0.5 μg/mL, a dose interval of at least 5 days can be achieved for DOX‐h‐LA, whereas p.o. and i.v. dosing of DOX‐h may only last 11 and 15 h, respectively. Pigs were slaughtered on day 30 after this trial, and no visible remnants of the preparation were detected neither fibrosis was observed after a thorough macroscopic and histopathological analysis.     

Courses for horses: Rethinking the use of proton pump inhibitors in the treatment of equine gastric ulcer syndrome

Oral omeprazole has been the cornerstone of equine gastric ulcer syndrome (EGUS) treatment for nearly 20 years. However, approximately 15–30% of equine squamous gastric disease (ESGD) cases and 75% of equine glandular gastric disease (EGGD) cases fail to heal within current treatment guidelines. Recently, a number of factors that may affect the efficacy of oral omeprazole have been highlighted and the pharmacodynamics of a number of novel proton pump inhibitors (PPIs) have been described in the horse. The purpose of this article is to review the factors that affect oral omeprazole efficacy, with the goal of maximising therapeutic response, and the novel PPIs recently described.     

Nephrotoxicity in Goats Caused by Dosing with a Water Extract from the Stems of Narthecium Ossifragum Plants

Seven goats were given a single dose of an aqueous extract derived from 30 g (wet weight) of Narthecium ossifragum per kg liveweight. Their serum creatinine and urea concentrations increased to day 5 but then fell to normal by day 10. Serum magnesium increased to day 4 and decreased to normal by day 9. Their serum calcium concentration was lower than normal on days 4, 5 and 6. Histopathological examination of the kidneys of goats killed or found dead 2, 4, 6, 8, 11 or 16 days after dosing revealed tubular epithelial cell degeneration and necrosis. Regeneration of the tubular epithelium and signs of interstitial fibroplast proliferation and fibrosis could be seen in animals killed on days 8, 11, 16 and 42. No signs of liver damage were observed in 3 goats dosed with the insoluble plant material from 40 g (wet weight) Narthecium ossifragum per kg liveweight. The total dose was divided into three doses, which were given intraruminally within 7 h. The activities of aspartate aminotransferase, -glutamyl-transferase and glutamate dehydrogenase remained within the normal range in all 10 goats after dosing.     

Evaluation of a glutamine-containing oral rehydration solution for the treatment of calf diarrhoea using an Escherichia coli model

A high-calorie oral rehydration solution (ORS) with glutamine (n=11) was more effective in correcting plasma, extracellular fluid and blood volume than solutions without (one WHO-type solution, n=6, and two high-glucose but glutamine-free solutions, n=7, n=12). It was the only solution to improve plasma volume significantly within 48 h and sustain the improvement throughout treatment; similarly, it was the only solution to correct packed-cell volume within 48 h and sustain the benefit to the end of treatment. At the end of treatment, the glutamine-treated calves were the only ones to avoid a significant weight loss compared with their pre-diarrhoeic values. The crucial difference between this solution and those used with glutamine previously is that it gave significant nutritional support whereas WHO type solutions did not. It also had more favourable effects on hyponatraemia and metabolic acidosis than a standard ORS. Use of a high-calorie ORS for 4 days (rather than 2 days of 50:50 admixture with milk replacer) brought additional beneficial effects on blood glucose and body weight.