Development of a two-step nested duplex PCR assay for the rapid detection of Brachyspira pilosicoli and Brachyspira intermedia in chicken faeces

Avian intestinal spirochaetosis (AIS) is an infection of the caeca and/or colo-rectum of laying and meat breeder hens caused by anaerobic intestinal spirochaetes of the genus Brachyspira. AIS can result in a variety of symptoms, including delayed and/or reduced egg production, and increased faecal water content. The two most commonly reported Brachyspira species involved in AIS are Brachyspira pilosicoli and Brachyspira intermedia, and their detection and identification can be difficult and time consuming. In the current study a two-step nested duplex PCR (2S-N-D-PCR) was developed for the detection of these two species, using DNA extracted from washed chicken faeces. In the first step, a duplex PCR (D-PCR) amplifying Brachyspira genus-specific portions of the 16S rRNA and NADH oxidase (nox) genes was undertaken on the washed faeces. In the second step, a nested D-PCR was used that amplified species-specific portions of the 16S rRNA gene of B. pilosicoli and the nox gene of B. intermedia from the amplicons produced in the first step. The 2S-N-D-PCR was rapid and specific, and could be used to detect approximately 10(3) cells of each spirochaete species per gram of washed faeces. When tested on 882 chicken faecal samples from infected flocks, it detected 4-5% more positive faecal samples than did the standard method of selective anaerobic culture followed by individual species-specific PCR assays conducted on the growth on the primary plate. The application of this new technique should improve diagnostic capacity, and facilitate further studies on AIS.     

Pharmacokinetics of ketoprofen in the green iguana (Iguana iguana) following single intravenous and intramuscular injections.

The nonsteroidal antiinflammatory drug ketoprofen (KTP) is a commonly used antiinflammatory and analgesic agent in reptile medicine, but no studies documenting its pharmacokinetics in this species have been published. Ketoprofen was administered as a racemic mixture to green iguanas (Iguana iguana) intravenously (i.v.) and intramuscularly (i.m.) at 2 mg/kg. Pharmacokinetic analyses were performed and indicated that ketoprofen in iguanas administered by the intravenous route has a classical two-compartmental distribution pattern, a slow clearance (67 ml/ kg/hr) and a long terminal half-life (31 hr) compared to ketoprofen studies reported in mammals. When delivered by the intramuscular route, bioavailability was 78%. These data indicate the daily dosing that is generally recommended for reptile patients, as an extrapolation from mammalian data, may be more frequent than necessary.     

A commercial trial evaluating three open water sources for farmed ducks: effects on health and production.

1. The present study investigated the effects on Pekin duck health and production of three open water resources in a commercial setting. The three resources were: narrow troughs (15?cm wide and 8?cm deep), intermediate troughs (20?cm wide and 12?cm deep) and wide troughs (50?cm wide and 8?cm deep). A total of 23 flocks of ducks were investigated, with an average size of 4540?±?680 individuals and a final stocking density always under 17?kg/m².

2. The health of the birds was considered to be of a good standard overall. The only health difference among the resources was that the foot condition of ducks provided with wide troughs was worse than those with intermediate troughs, whilst the foot condition of birds with narrow troughs was intermediate. The open water resources did not differ in their effects on the production measures assessed.

3. Properly managed open water did not have any major negative effects on the health or production of commercial Pekin ducks. Wide, intermediate and narrow troughs did not differ substantially in measures of health or productivity.     

Periovulatory endocrinology and oocyte maturation in unmated mature blue fox vixens (Alopex lagopus)

Nine of 10 mature blue fox vixens (Alopex lagopus) in spontaneous oestrus ovulated approximately 2 days after the preovulatory increase in luteinizing hormone (LH). Plasma concentrations of follicle-stimulating hormone and progesterone increased simultaneously with the LH peak, whereas oestradiol-17 beta peaked 1 day previously. In the tenth vixen, an LH peak was not observed, and neither visible follicles nor corpora lutea were found in the ovaries 6 days after peak vaginal electrical resistance. Eggs were ovulated as primary oocytes, but oocyte maturation was initiated within the day of ovulation (2 days after the LH peak). Within the next 2 days (3-4 days after the LH peak) the first polar body was extruded, and the cumulus mass was completely dissociated from the zona pellucida. The interval between the preovulatory LH peak and initiation of the final oocyte maturation is thus considerably longer in the blue fox than for example in the cow (48-72 h compared with 9-12 h). This suggests that the relationship between these two events is somewhat different in the blue fox.     

Large intestinal capacity, retention times, and turnover rates of particulate ingesta associated with extensive large-colon resection in horses

Fecal excretion of a particulate marker, ytterbium (Yb), was evaluated in 9 horses before surgery and 3 weeks, 3 months, and 6 months (4 trials) after sham-operation (group 1; n = 3) or extensive large colon resection (group 2; n = 6). Fecal excretion curves of total Yb excretion, loge Yb excretion, % Yb excretion, loge % Yb excretion, and cumulative % Yb excretion were evaluated, and kinetic analysis was performed on the loge Yb excretion curves to detect mixing pools and to calculate the fractional rate of particulate passage, turnover rate, and pool size. Calculations were performed to determined transit time, mean overall retention time, adjusted mean retention time, peak time, and disappearance time. Values were statistically analyzed to determine differences between groups and among trials (P less than 0.05). Group-2 horses had significantly shorter transit, peak, and mean overall retention times, compared with preoperative values and with values for group-1 horses. Two mixing pools were identified: a slower emptying pool of 5.7% hour-1 (k1) and a faster emptying pool of 12.3% hour-1 (k2). The rate of passage from the first pool (k1) was not altered by colon resection, and was interpreted as being most influenced by the cecum. In further support of this interpretation, the capacity of the k1 pool approximated the capacity of the cecum (17 L). The capacity of the k1 pool significantly expanded by 6 months in the resected horses. the rate of passage from the second pool (k2) significantly increased initially after colon resection (3 weeks and 3 months), but returned to preoperative values by 6 months.(ABSTRACT TRUNCATED AT 250 WORDS)     

In vitro reactivity of digital arteries and veins to vasoconstrictive mediators in healthy horses and in horses with early laminitis

The in vitro reactivity of vasoconstrictive mediators that are implicated in acute laminitis was determined in palmar and plantar digital arteries and veins obtained from healthy horses and in palmar digital vessels of horses with early laminitis (Obel grade I). To obtain baseline reactivity data, 3 experiments were conducted, using healthy horses: (1) the reactivity of palmar and plantar digital arteries and veins to angiotensin II, norepinephrine, and 5-hydroxytryptamine (serotonin) were compared; (2) the direct effects of bacterial endotoxin on vascular reactivity were assessed; and (3) the reactivity of palmar digital arteries and veins to angiotensin II, norepinephrine, prostaglandin F2 alpha (PGF2 alpha), serotonin, and a thromboxane-endoperoxide analog (U46619) were determined. The vascular reactivity of these same 5 vasoconstrictors then was determined in horses with early laminitis and was compared with data from healthy (control) horses. Obel grade-I laminitis was experimentally induced in horses, using carbohydrate overload. Dose responses were conducted for each agent at concentrations between 10(-8)M and 10(-4)M. The potency of a drug was defined as the mean effective concentration necessary to induce 50% of maximal contraction (EC50). There were no differences in EC50 concentrations and in maximal contractions between forelimb and hind limb arteries and veins for angiotensin II, norepinephrine, and serotonin. Incubation with endotoxin had no effect on the reactivity of arteries and veins to angiotensin II, norepinephrine, and serotonin.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of weight loading on the coronary band interstitial fluid pressure in horses

Interstitial fluid pressures, as a possible function of limb load, were measured at 2 sites within the digital coronary dermis of both cranial digits in 10 standing horses. Fluid pressure changes and digital load measurements were simultaneously detected and recorded by use of, respectively, modified wick-in-needle and force plate transducers coupled to a microcomputer. Mean pressures, recorded at limb loads between 50 and 80 kg, were 2.29 +/- 3.17 mm of Hg at the toe and 2.49 +/- 5.91 mm of Hg at the heel. Mean pressures, recorded between 150 and 180 kg, were 5.01 +/- 5.23 mm of Hg at the toe and 1.28 +/- 7.69 mm of Hg at the heel. These data indicate that, in the static limb, no statistically significant change in interstitial fluid pressure occurs at loads up to 180 kg.     

牛子宫复旧过程中胶原蛋白和肌浆球蛋白降解规律的研究

本实验监测8头围产期奶牛外周血浆中胶原蛋白和肌浆球蛋白的代谢产物——羟基脯氨酸(HP)和3甲基组氨酸(3MH)的含量,借以找出这两种蛋白质的代谢规律和子宫复旧的关系。结果表明:荷兰黑白花牛和娟珊牛血浆中HP的含量和子宫角直径分别呈显著及极显著正相关(r=0.41,P<0.05和r=0.57 P<0.01),而3MH分别呈极显著和显著正相关(r=0.52 P<0.01和r=0.39(P<0.05)。此外,黑白花牛血浆中3MH的含量极显著高于娟珊牛(P<0.001)。两个品种牛子宫复旧的平均时间,黑白花牛为37.4±3.1天,娟珊牛32.3±2.1天。结果说明牛产后子宫体积的缩小与子宫肌肉的胶原蛋白和肌浆球蛋白降解成氨基酸排出体外有关,子宫复旧的速度可能决定于这两种蛋白质的降解速度。