Feline vaccine‐associated sarcomagenesis: Is there an inflammation‐independent role for aluminium?

Aluminium has been found in feline vaccine‐associated sarcomas. In this study, we investigated the potential for aluminium to contribute directly to tumourigenesis. Our results indicated that an aluminium hydroxide adjuvant preparation was cytotoxic and mutagenic in human‐Chinese hamster ovary (CHO) hybrid cells in vitro. Moreover, CHO cells deficient in DNA double strand break (DSB), but not single‐strand break (SSB), repair, were particularly sensitive to aluminium exposure compared with repair proficient cells, suggesting that aluminium is associated with DSBs. In contrast to CHO cells, primary feline skin fibroblasts were resistant to the cytotoxic effects of aluminium compounds and exposure to an aluminium chloride salt promoted cell growth and cell cycle progression at concentrations much less than those measured in particular feline rabies vaccines. These findings suggest that aluminium exposure may contribute, theoretically, to both initiation and promotion of tumours in the absence of an inflammatory response.     

Risk Factors for the Development of Hypokalemia in Neonatal Diarrheic Calves

Background

Neonatal diarrheic calves have a clear negative potassium balance because of intestinal losses and decreased milk intake but in the presence of acidemia, they usually show normokalemic or hyperkalemic plasma concentrations.

Objectives

To assess whether marked hypokalemia occurs in response to the correction of acidemia and dehydration and to identify factors that are associated with this condition.

Animals

Eighty‐three calves with a clinical diagnosis of neonatal diarrhea.

Methods

Prospective cohort study. Calves were treated according to a clinical protocol using an oral electrolyte solution and commercially available packages of 8.4% sodium bicarbonate, 0.9% saline and 40% dextrose infusion solutions.

Results

The proportion of hypokalemic calves after 24 hours of treatment (19.3%) was twice as great as it was on admission to the hospital. Plasma K⁺ after 24 hours of treatment was not significantly correlated to venous blood pH values at the same time but positively correlated to venous blood pH values on admission (r = 0.51, P < .001). Base excess on admission (Odds ratio [OR] = 0.81, 95% confidence interval [CI] = 0.70–0.94), duration of diarrhea (OR = 1.37, 95% CI = 1.05–1.80), milk intake during hospitalization (OR = 0.54, 95% CI = 0.37–0.79) and plasma sodium concentrations after 24 hours (OR = 1.12, 95% CI = 1.01–1.25) were identified to be independently associated (P < .05) with a hypokalemic state after 24 hours of treatment.

Conclusions and Clinical Importance

Findings of this study suggest that marked depletion of body potassium stores is evident in diarrheic calves that suffered from marked metabolic acidosis, have a low milk intake and a long history of diarrhea.     

Citric acid extraction—An underestimated method in forest nutrition?

Since recent studies reveal citric acid to be favorable for estimating plant‐available P in soils, we investigated if it can also be used for assessing other nutrients. According to our results, it provides stronger correlations with the tree nutrition for Mg (beech, spruce), Ca+K (beech) and Fe (spruce) than the standard methods for determining exchangeable cations. Thus, when estimating plant‐available P by citric acid‐extraction, these cations should be additionally measured in ICP analysis.     

The α2A‐adrenoceptor subtype plays a key role in the analgesic and sedative effects of xylazine

Xylazine, the classical α₂‐adrenoceptor (α₂‐AR) agonist, is still used as an analgesic and sedative in veterinary medicine, despite its low potency and affinity for α₂‐ARs. Previous pharmacological studies suggested that the α_2A‐AR subtype plays a role in mediating the clinical effects of xylazine; however, these studies were hampered by the poor subtype‐selectivity of the antagonists used and a lack of knowledge of their bioavailability in vivo. Here, we attempted to elucidate the role of the α_2A‐AR subtype in mediating the clinical effects of xylazine by comparing the analgesic and sedative effects of this drug in wild‐type mice with those in α_2A‐AR functional knockout mice using the hot‐plate and open field tests, respectively. Hippocampal noradrenaline turnover in both mice was also measured to evaluate the contribution of α_2A‐AR subtype to the inhibitory effect of xylazine on presynaptic noradrenaline release. In wild‐type mice, xylazine (10 or 30 mg/kg) increased the hot‐plate latency. Furthermore, xylazine (3 or 10 mg/kg) inhibited the open field locomotor activity and decreased hippocampal noradrenaline turnover. By contrast, all of these effects were abolished in α_2A‐AR functional knockout mice. These results indicate that the α_2A‐AR subtype is mainly responsible for the clinical effects of xylazine.     

Clinical evaluation of the anaesthetic sparing effect of brachial plexus block in cats

ObjectiveTo evaluate the isoflurane sparing effect and the post-surgical analgesia provided by a brachial plexus block (BPB) in cats undergoing distal thoracic limb surgery.Study designProspective randomized blinded clinical study.AnimalsTwenty client-owned cats.MethodsCats were assigned to receive either no BPB (group NB) or a nerve stimulator guided BPB (group BPB) using lidocaine (3.6 mg kg^?1) and bupivacaine (1.2 mg kg^?1). Pre-medication consisted of midazolam and ketamine intravenously (IV). Anaesthesia was induced with propofol IV to effect and maintained with isoflurane delivered in oxygen and a continuous rate infusion of fentanyl (2 μg kg^?1 hour^?1). End-tidal isoflurane concentration (Fe′ISO) was adjusted every 3 minutes guided by changes in cardiorespiratory parameters and reflexes present, to maintain a stable depth of anaesthesia. Five time points were chosen to record all parameters and compare values between groups. Recovery and post-operative pain assessment were performed using a visual analogue scale (VAS) at 15 and 45 minutes after extubation and thereafter at hourly intervals until 5 hours after placement of the BPB.ResultsNo clinically significant differences were seen for heart rate, respiratory rate and non-invasive blood pressure between groups. Mean Fe′ISO was significantly lower in group BPB compared with group NB at all time points. In group NB, all intraoperative measurements of Fe′ISO were significantly higher compared with baseline (3 minutes before start of surgery) measurements. During recovery, VAS scores for group BPB were significantly lower than for group NB. Additional analgesics were needed in all cats within the study period.Conclusion and Clinical relevanceIn cats undergoing orthopaedic surgery of the thoracic limb, BPB reduced intra-operative isoflurane requirement and pain during the early post-operative period when compared with procedures without a BPB. BPB is a useful adjunct to anaesthesia in such cases.     

Effects of pistachio by‐products on digestibility,milk production,milk fatty acid profile and blood metabolites in Saanen dairy goats

The objective of this study was to investigate the effects of pistachio by‐products (PBP) on nutrient digestibility, blood metabolites and milk fatty acid (FA) profile in Saanen dairy goats. Nine multiparous lactating Saanen goats (on day 90 post‐partum, 45 ± 2/kg BW) were randomly assigned to a 3 × 3 Latin square design with three treatment diets: 1) control diet (alfalfa hay based), 2) 32% PBP and 3) 32% PBP + polyethylene glycol (PEG‐4000; 1 g/kg dry matter). Each period lasted 21 days, including 14 day for treatment adaptation and 7 day for data collection. Pistachio by‐products significantly decreased (p < 0.01) crude protein (CP) digestibility compared with the control diet (64.4% vs. 58.7%), but PEG addition did not differ for CP digestibility of goats fed 32% PBP + PEG and those fed the two other diets. The digestibility of NDF tended (p = 0.06) to decrease for goats fed PBP compared with those fed the control diet. Yields of milk and 4% fat‐corrected milk were not affected by dietary treatments. Compared with the control diet, PBP supplementation appreciably changed the proportions of almost all the milk FA measured; the main effects were decreases (p < 0.01) in FA from 8:0 to 16:0 and increases (p < 0.01) proportions of cis‐9, trans‐11 18:2 and trans‐11 18:1, monounsaturated FA, polyunsaturated FA and long‐chain FA. The saturated FA, short‐chain FA and medium‐chain FA proportions were lower (p < 0.01) in goats fed the two PBP supplemented diet than in those fed the control diet and PEG addition led to intermediate proportions of saturated FA, unsaturated and monounsaturated FA. Inclusion of PBP in the diet decreased (p < 0.01) plasma concentrations of glucose and urea nitrogen compared with the control diet. It was concluded that PBP can be used as forage in the diet of dairy goats without interfering with milk yield. Inclusion of 32% PBP in the diet of dairy goats had beneficial effects on milk FA profile but PEG addition to PBP did not contribute to enhance further milk FA profile.     

Impressions and purchasing intentions of Japanese consumers regarding pork produced by ‘Ecofeed,’ a trademark of food‐waste or food co‐product animal feed certified by the Japanese government

Impressions and purchasing intentions of Japanese consumers regarding pork produced by ‘Ecofeed’, a trademark of food‐waste or co‐product animal feeds certified by the Japanese government, were investigated by a questionnaire on the Internet. ‘Ecofeed’ did not elicit specific impressions as compared to domestic, imported, Kurobuta (in Japan), and specific pathogen‐free (SPF) pork. Purchasing intent for ‘Ecofeed’ pork was the second lowest of the five pork products. Knowledge and purchasing experience regarding ‘Ecofeed’ pork was the lowest of the five pork products. Respondents were classified into four categories according to their impressions of ‘Ecofeed’ pork. The largest category of respondents did not have any specific impression of ‘Ecofeed’ pork and had little knowledge of pork farming. A category that had a positive impression for ‘Ecofeed’ pork had high knowledge of the pork farming system. In order to establish ‘Ecofeed’ pork in Japan, our results suggest that information disclosure and education about ‘Ecofeed’, its certification system, environmental benefits and the current self‐efficiency ratio of animal feed, are needed.     

Ralstonia solanacearum virulence in tomato seedlings inoculated by leaf clipping

Ralstonia solanacearum is a phytopathogenic bacterium that colonizes the xylem vessels of host plants leading to a lethal wilt disease. Although several studies have investigated the virulence of R. solanacearum on adult host plants, infection studies of this pathogen on the seedling stages of hosts are less common. In a preliminary observation, inoculation of R. solanacearum F1C1 on 6‐ to 7‐day‐old tomato seedlings by a simple leaf‐clip strategy resulted in a lethal pathogenic condition in seedlings that eventually killed these seedlings within a week post‐inoculation. This prompted testing of the effect of this inoculation technique in seedlings from different cultivars of tomato and similar results were obtained. Colonization and spread of the bacteria throughout the infected seedlings was demonstrated using gus‐tagged R. solanacearum F1C1. The same method of inoculating tomato seedlings was used with R. solanacearum GMI1000 and independent mutants of R. solanacearum GMI1000, deficient in the virulence genes hrpB, hrpG, phcA and gspD. Wildtype R. solanacearum GMI1000 was found to be virulent on tomato seedlings, whereas the mutants were found to be non‐virulent. This leaf‐clip technique, for inoculation of tomato seedlings, has the potential to be a valuable approach, saving time, space, labour and costs.     

Nitric oxide impacts bovine sperm capacitation in a cGMP‐dependent and cGMP‐independent manner

We aimed to elucidate whether NO acts in in vitro sperm capacitation in bovine via cGMP/PKG1 pathway. For this, cryopreserved bovine sperm were capacitated in vitro with 20 µg/ml heparin (Control) plus treatments: 1 mM L‐arginine (L‐arg, NO precursor), 50 µM Rp‐8‐Bromo‐β‐phenyl‐1,N²‐ethenoguanosine‐3′,5′‐cyclic monophosphorothioate (Rp‐8‐Br‐cGMPS, selective inhibitor of the binding site for cGMP in PKG1), 1 mM 2‐Phenyl‐4,4,5,5‐tetramethylimidazoline‐1‐oxyl 3‐oxide (PTIO, NO scavenger), and the combinations of L‐arg + RP‐8‐Br‐cGMPS and L‐arg + PTIO. Sperm motility and vigour were determined by phase‐contrast microscopy, capacitation status by chlortetracycline staining, and the intracellular concentration of cGMP was measured by ELISA. Data were subjected to analysis of variance and means compared with SNK test at 5% probability. Motility and vigour were lower in sperm treated with PTIO when compared to Control and other treatments (p < .05). The L‐arg treatment showed the highest percentage of capacitated sperm when compared to the Control and other treatments (Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO) (69.8 ± 3.4%, 51.2 ± 3.0, 51.1 ± 2.1, 51.2 ± 3.0 and 45.5 ± 2.7, respectively) (p < .05). The capacitation ratio (%) was lower in treatments with Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO, respectively (p < .05). Lastly, cGMP concentration (pmol/ml) was lower in PTIO and L‐arg + PTIO (1.3 ± 0.3 and 1.6 ± 0.4) and was higher in Rp‐8‐Br‐cGMPS and L‐arg + Rp‐8‐Br‐cGMPS (3.7 ± 0.4 and 4.0 ± 0.5) treatments. We showed that during in vitro capacitation of cattle: (a) NO influences sperm motility and vigour; (b) NO is associated with cGMP synthesis through two independent pathways and (c) the cGMP/PKG1 pathway has a partial role in sperm capacitation and does not involve the L‐arg/NO.