Adenosine A2A receptor agonists inhibit lipopolysaccharide-induced production of tumor necrosis factor-alpha by equine monocytes

Adenosine is an endogenous nucleoside that regulates many physiological processes by activating one or more adenosine receptor subtypes, namely A1, A2A, A2B and A3. The results of previous studies indicate that adenosine analogues inhibit lipopolysaccharide (LPS)-induced production of reactive oxygen species (ROS) by equine neutrophils primarily through activation of A2A receptors. Because peripheral blood monocytes produce cytokines that are responsible for many of the deleterious effects of LPS, the current study was performed to evaluate the effects of an array of novel adenosine receptor agonists on LPS-induced production of tumor necrosis factor-alpha (TNF-alpha), and to assess the selectively of these agonists for equine adenosine A2A over the A1 receptor. Radioligand binding studies performed with equine tissues expressing adenosine A1 and A2A receptor subtypes yielded a rank order of affinity for the equine A2A receptor of ATL307>ATL309 approximately ATL310 approximately ATL313>ATL202 approximately ATL361 approximately ATL376>ATL372>CGS21680>NECA. Co-incubation of equine peripheral blood monocytes with LPS and these agonists resulted in inhibition of TNF-alpha production with a rank order of potency that strongly correlated with their binding affinities for equine adenosine A2A receptors. Results of experiments performed with one of the adenosine receptor agonists (ATL313) and selective adenosine receptor antagonists confirmed that inhibition of LPS-induced production of TNF-alpha occurred via stimulation of A2A receptors. Although incubation of monocytes with IB-MECA, a compound purported to act as an adenosine A3 receptor agonist, reduced LPS-induced TNF-alpha production, this effect of IB-MECA was inhibited by the A2A selective antagonist ZM241385 but not by the A3 receptor antagonist MRS1220. These results indicate that the adenosine receptor subtype responsible for regulation of LPS-induced cytokine production by equine monocytes is the A2A receptor. To address the signal transduction mechanism responsible for the anti-inflammatory effects of ATL313 in equine monocytes, production of cAMP was compared in the presence and absence of either the adenosine A2A receptor antagonist ZM241385 or the adenosine A2B receptor antagonist MRS1706. In the absence of the antagonists, ATL313 increased production of cAMP; ZM241385 inhibited this effect of ATL313, whereas MRS1706 did not. Furthermore, incubation of monocytes with either the stable analogue of cAMP, dibutyryl cAMP, or forskolin, an activator of adenylyl cyclase, also inhibited LPS-induced production of TNF-alpha production by equine monocytes. Collectively, the results of the current study indicate that adenosine analogues inhibit LPS-induced production of TNF-alpha by equine monocytes primarily via activation of adenosine A2A receptors and do so in a cAMP-dependent manner. The results of this study indicate that stable adenosine analogues that are selective for adenosine A2A receptors may be suitable for development as anti-inflammatory drugs in horses.     

The effect of omeprazole paste on intragastric pH in clinically ill neonatal foals

REASON FOR PERFORMING STUDY: Administration of omeprazole paste per os to healthy neonatal foals has been shown to effectively increase intragastric pH, but has not been evaluated in sick neonatal foals. OBJECTIVES: To determine the effect of orally administered omeprazole paste on intragastric pH in clinically ill neonatal foals requiring nasogastric intubation. METHODS: Intragastric pH was measured continuously for 24 h using an indwelling electrode and continuous data recording system in hospitalised neonatal foals age < or =2 days. Intragastric pH was measured for 12 h prior to (pretreatment period) and 12 h following (post treatment period) treatment with omeprazole paste (4 mg/kg bwt per os). All foals displayed periods of acidity (pH <4) prior to treatment. Statistical analysis compared pre- and post treatment mean and median intragastric pH, and percentage of time below pH 4. RESULTS: Eight foals were evaluated age 1-3 days, a gestational age of at least 320 days or reported to be full term. The mean (3.19 +/- 1.50 vs. 6.20 +/- 0.93) and median (4.6 +/- 1.7 vs. 6.86 +/- 0.89) pH were significantly higher and the percentage of time below pH 4 (32.25 vs. 1.1%) was significantly lower in the post treatment compared to the pretreatment period. CONCLUSION: Omeprazole paste effectively increases intragastric pH in clinically ill neonatal foals after one dose at 4 mg/kg bwt orally.     

The effects of increasing sodium chloride concentration on Mycoplasma gallisepticum vaccine survival in solution

Lyophilized Mycoplasma gallisepticum (MG) vaccines are generally rehydrated and diluted with distilled or chlorine-free water as per manufacturer recommendations. However, as mycoplasma species lack a cell wall, this can lead to decreased viability of live vaccine during administration. The ability of phosphate-buffered saline (PBS) to prevent losses in live vaccine viability was examined. It was shown that a concentration of 1 x PBS prevented the two-fourfold decrease in MG viability seen when the vaccines were diluted with water alone.     

Pathological fractures secondary to metaphyseal osteopathy in a dog.

A four-month-old German Shepherd Dog (GSD) was admitted with bilateral non-weight bearing hindlimb lameness and pyrexia. A diagnosis of bilateral distal femoral fractures secondary to metaphyseal osteopathy was made on the basis of clinical signs, radiography and biopsy. The dog returned to normal function following conservative management. As far as the authors are aware there are no previous reports of pathological fractures secondary to this condition.     

Spoligotype diversity of Mycobacterium bovis and Mycobacterium caprae animal isolates

The genetic diversity of 283 Mycobacterium bovis (M. bovis) and 10 Mycobacterium caprae (M. caprae) strains, isolated between 2002 and 2007 from cattle, goat, red deer and wild boar from six different geographical regions of Portugal was investigated by spoligotyping. The technique showed a good discriminatory power (Hunter-Gaston Index, h=0.9) for the strains, revealing 29 different patterns. One pattern (SB0121) was clearly predominant, accounting for 26.3% of the isolates; ten patterns, representing 20.7% of the isolates, had never been reported previously. Multiple spoligotypes were detected in thirteen cattle and one goat herd, most of which were found in beef cattle and extensive management regions, suggesting different infection sources. With the exception of two spoligotypes, those in wildlife species were also found in domestic species.     

Ocular distribution and toxicity of intravitreal injection of triamcinolone acetonide in normal equine eyes

Objective  To determine ocular distribution and toxicity of a single injection of intravitreal triamcinolone acetonide (TA) in normal horses.
Animals studied  Six adult horses, donated to North Carolina State University.
Procedures  Six horses were injected intravitreally with either 10, 20, or 40 mg ( n  = 2 each) of TA. The opposite eye of each horse was injected with balanced salt solution (BSS). Ocular toxicity was assessed by biomicroscopy, tonometry, indirect ophthalmoscopy, and electroretinogram. Aqueous humor (AH), vitreous humor (VH), and plasma samples were collected. Horses were euthanized 7 or 21 days after injection and eyes enucleated for histopathology. TA concentrations in AH, VH, and plasma were measured by HPLC.
Results  Three control eyes and one TA eye developed inflammation after injection or collection of AH. Positive bacterial cultures ( Corynebacterium spp., Staphylococcus spp., and Streptococcus spp.) were obtained from three of these eyes. Other than transient corneal edema in TA injected eyes, which resolved by 7 days after injection, no other changes were observed. TA crystals were visible within the vitreous body. No evidence of TA toxic effect was noted on histopathology. TA was detected in all AH and VH samples from treated eyes following injection. Drug was not detected in the plasma.
Conclusions  There was no evidence of overt toxicity from intravitreal TA in normal horses and a single intravitreal injection resulted in TA ocular levels for 21 days. However, the risk for bacterial infections with intravitreal injection or anterior chamber aspirations in horses is high. Use of topical and systemic antibiotics after injection is recommended.     

Feeding Acacia etbaica and Dichrostachys cinerea fruits to smallholder goats in northern Ethiopia improves their performance during the dry season

Acacia etbaica Schweinf. and Dichrostachys cinerea (L.) Wight et Arn. are among the common woody browse plants that naturally grow in many arid and semi-arid rangelands in sub Saharan Africa. An experiment was conducted to compare the voluntary dry matter intake, body weight gain, nitrogen balance, carcass composition and sensory attributes of goats supplemented with different levels of A. etbaica and D. cinerea fruits. Average initial body weight of the goats was 20.4 kg (± 0.8SE) and received one of the following fruit supplementation regimes for 120 days (on body weight basis, n = 4 goats per group): (1) Control, no supplement; (2) 0.5% of A. etbaica; (3) 0.5% of D. cinerea; (4) 1.0% of A. etbaica; (5) 1.0% of D. cinerea; (6) 1.5% of A. etbaica; (7) 1.5% of D. cinerea. All groups were allowed to graze/browse during the day. Compared to A. etbaica, D. cinerea fruits contained higher amounts of crude protein (182 vs 135 g/kg DM), metabolizable energy (11 vs 8.4 MJ/kg DM), and in vitro dry matter digestibility coefficient (0.7 vs 0.6). The concentrations of acid detergent fiber (ADF), sulphuric acid solubulized lignin, and tannin (soluble and condensed) were lower (P < 0.05) in D. cinerea than in A. etbaica fruits. A. etbaica fruits, however, contained higher amounts of Na, K, Fe, and Zn concentrations than D. cinerea fruits. Dry matter intake was markedly higher in supplemented groups than in the control group without supplement. Similarly, body weight gain in the control group was negative and lower (P < 0.05) than any of the supplemented groups. During the course of the experimental period (120 days) the group placed in the 1.5% D. cinerea fruit diet gained 2.6 kg whereas the control group lost 2.0 kg. Thus, the difference between these two groups was 4.6 kg, which suggests about 22.5% increment of the goats' initial body weight. N retention was negative for the control group and varied little among the remaining treatment groups. Dressing percent increased with increased level of either type of fruit supplement, the highest (51.8%) being achieved at the 1.5% D. cinerea level. There was no difference in bone tissue across treatment groups. D. cinerea fruit intake also resulted in juicy/watery flavor and tender carcass. D. cinerea fruits appeared to have no adverse effects when included at the highest level (1.5%) and could be collected and stored as dry season supplement to smallholder goats. This study shows that poor farmers can increase goat performance substantially through supplement feeding with wild fruits. Areas in Ethiopia originally set aside for provision of ecological services and biodiversity protection can also be used for wild fruit production and feeding of small stock. The effect of increasing D. cinerea fruits above the 1.5% body weight level should be a focus of future investigation. Further work on deactivation mechanism of tannins is required for A. etbaica fruits included at levels higher than 1.0%.     

Naked oats: metabolisable energy yield from a range of varieties in broilers, cockerels and turkeys

1. Naked oats belong to the same species as 'common oats', Avena sativa, but have a non-lignified husk which readily becomes detached during harvesting. The absence of the indigestible husk can be predicted to give an increased metabolisable energy (ME) content for poultry. 2. Measurements of true metabolisable energy (TME(N)) were performed on 3-week-old broiler chicks (Cobb males), adult cockerels (ISA Brown, greater than one year old) and 7-week-old turkeys (BUT T8 males). The measurements were repeated in 2000, 2001 and 2002, with some measurements on a subset of varieties in 2004. 3. High-oil naked oat lines yielded 12% more energy (TME(N)) than wheat. Naked oats, excluding the experimental high-oil lines, yielded 8.5% more energy than simultaneously assayed wheat samples. 4. In samples from the 2004 harvest, conventional oats gave TME(N) values about 13% lower than those of wheat. 5. The addition of beta-glucanase produced an increase of about 4% in the apparent metabolisable energy (AME) of oats for broiler chickens. This effect was associated with a 70% decrease in the jejunal viscosity of broilers receiving a 500 g/kg naked-oat diet. 6. The oil content of naked oats was about 5 times greater than that of wheat, with the high-oil lines rising to more than 6 times greater. Naked oats had a lower starch content than wheat but not sufficiently lower to negate the energy benefits of the higher oil content. The crude protein (CP) contents of naked oats were similar to those of wheat, with the high-oil varieties tending to be higher in CP also.     

Dog-associated risk factors for human plague

Plague is a rare but often fatal zoonosis endemic to the western United States. Previous studies have identified contact with pets as a potential risk factor for infection. We conducted a matched case-control study to better define the risks associated with pets at both the household and individual levels. Using a written questionnaire, we surveyed nine surviving plague patients, 12 household members of these patients, and 30 age- and neighbourhood-matched controls about household and individual exposures. Overall, 79% of households had at least one dog, 59% had at least one cat and 33% used flea control, with no significant differences between case and control households. Four (44%) case households reported having a sick dog versus no (0%) control households [matched odds ratio, (mOR) 18.5, 95% confidence interval (CI) 2.3-infinity], and four (44%) patients reported sleeping in the same bed with a pet dog versus three (10%) controls (mOR 5.7, 95% CI 1.0-31.6). Within case households with multiple members, two (40%) of five patients slept with their dogs versus none (0%) of 12 healthy family members (P=0.13). The exposures to cats were not significant. Sleeping in the same bed as a pet dog remained significantly associated with infection in a multivariate logistic regression model (P=0.046). Our findings suggest that dogs may facilitate the transfer of fleas into the home and that activities with close extended contacts with dogs may increase the risk of plague infection.