Development of a technique for the in vivo assessment of flatulence in dogs.

OBJECTIVE: To develop a noninvasive method for the in vivo assessment of flatulence in dogs. ANIMALS: 8 adult dogs. PROCEDURE: Rectal gases were collected via a perforated tube held close to each dog's anus and attached to a monitoring pump fitted with a sensor that recorded hydrogen sulfide concentrations every 20 seconds. Patterns of flatulence were monitored for 14 hours after feeding on 4 days, and within- and between-dog variation was assessed over 4 hours on 4 consecutive days. Rate of hydrogen sulfide production (flatulence index) and frequency and number of emissions were evaluated as potential indicators of flatus characteristics. An odor judge assigned an odor rating to each flatulence episode, and the relationship between that rating and hydrogen sulfide concentration was determined. RESULTS: Flatulence patterns varied within and between dogs. Variation was most pronounced for flatulence index; mean coefficients of variance within dogs over time and between dogs on each day were 75 and 103%, respectively. Flatus with hydrogen sulfide concentrations > 1 parts per million could be detected by the odor judge, and severity of malodor was highly correlated with hydrogen sulfide concentration. Odor ratings were accurately predicted by use of the equation 1.51 X hydrogen sulfide concentration(0.28). CONCLUSIONS AND CLINICAL RELEVANCE: The technique described in this report appears to provide sensitive, reliable, and relevant data and will enable further studies of the factors that influence flatulence in dogs. Use of this technique also has the potential to aid in investigations of colonic physiology and pathology.     

Needle catheter duodenostomy: a technique for duodenal alimentation of birds

A technique for duodenal alimentation (needle catheter duodenostomy) of birds was developed, using the domestic pigeon (Columba livia) as the experimental model. A needle catheter was inserted into the descending duodenum of 5 pigeons and was secured to the body wall and dorsum of each bird. A liquid diet was administered daily (in equal amounts of 0, 4, 8, 12, and 16 hours) for 14 days without adverse effects. On day 15, the catheters were removed, and the birds immediately resumed normal consumption of a pigeon ration and water diet. Although 4 of the 5 birds had minor weight loss, dietary alterations probably could be used on an individual basis to alleviate this problem. After oral alimentation was resumed, the 5 birds exceeded their initial body weight within 7 days. Four weeks after catheter removal, positive-contrast radiographic evaluations indicated that the duodenum of each pigeon appeared normal. Needle catheter duodenostomy was a viable method of alimentation in the domestic pigeon. This technique should be applicable for other avian species requiring bypass of the upper gastrointestinal tract proximal to the region of catheter insertion in the duodenum.     

A technique for evaluating analgesia in conscious perching birds

OBJECTIVE: To develop a technique for objective assessment of modulation of nociperception in conscious perching birds. ANIMALS: 31 adult African grey parrots. PROCEDURE: Birds were randomly assigned to receive saline (0.9% NaCl) solution (n = 10), butorphanol tartrate (11), or buprenorphine hydrochloride (10), i.m. Birds were fitted with a surface electrode on the medial metatarsus of 1 leg. An electrical stimulus was delivered to the bird's foot through an aluminum surface on half of the perch. The alternate side of the perch delivered a noxious thermal stimulus. A withdrawal response to either stimulus was recorded when the bird lifted its foot or vigorously flinched its wings. RESULTS: Responses to thermal stimuli were extremely variable during baseline testing and after administration of drugs. Thus, significant differences were not detected after drug injection. In contrast, responses to an electrical stimulus were predictable with much less variation. CONCLUSIONS AND CLINICAL RELEVANCE: This method and device allowed for the reliable determination of withdrawal threshold in perching birds. Use of this technique for objective assessment of modulation of nociperception in conscious perching birds will enable assessment of analgesic drugs.     

PCR技术在鸟类性别鉴定上的应用

根据鸟类性染色体连锁基因EE0．6在Z、W染色体上长度的不同，通过选择6对不同的引物，同时设置4种不同的反应条件，分别采用PCR技术对丹顶鹤、黑鹳、琉璃金刚鹦鹉、绯红金刚鹦鹉、鞭苔鹞鹚、戴冕鹤、火烈鸟的Z、W染色体上的EE0．6基因进行了特异性扩增。结果显示，某些引物组合能从雄鸟样品中扩增出1条片段（EE0．6Z，190bp），而雌性鸟样品则能扩增出2条片段（EE0．6W和EE0．6Z，分别为250bp和190bp），但4组引物对6种鸟类EE0．6基因的扩增效果不同。证实，利用PCR技术能对鸟类进行性别鉴定。     

Serum from dogs Infected with Babesia gibsoni inhibits maturation of reticulocytes and erythrocyte 5'-nucleotidase activity in vitro

Erythrocyte 5'-nucleotidase is thought to be involved in the maturation of erythrocytes. In the present study, in vitro incubation of canine erythrocytes demonstrated that significant inhibition of 5'-nucleotidase activity occurred in the presence of serum from dogs infected with Babesia gibsoni, when the enzyme was assayed with cytidine 5'-monophosphate (5'-CMP) and inosine 5'-monophosphate (5'-IMP) as substrates. The multiplication of B. gibsoni in in vitro culture also resulted in a significant decrease in the enzyme activity of erythrocytes in the culture. Furthermore, the infected serum and 5'-CMP retarded the maturation of canine reticulocytes in vitro. These results suggested that nucleotides such as 5'-CMP and 5'-IMP might accumulate in young erythrocytes and/or serum in dogs infected with B. gibsoni as a result of decreased activity of erythrocyte 5'-nucleotidase, resulting in the delayed maturation of reticulocytes.     

Potassium chloride as a euthanasia agent in psittacine birds: Clinical aspects and consequences for histopathologic assessment

Twelve parrots anesthetized with isoflurane were euthanized intravenously (IV) with 3 or 10 mEq/kg body weight (BW) of potassium chloride (KCl) resulting in ventricular asystole at 68.0 s and 32.8 s, respectively. Mild vocalization (1/6 birds, 3 mEq/kg BW) and involuntary muscle tremors (5/6 birds, 10 mEq/kg BW) were noted. Unlike barbiturates or T-61 no histologic artefacts resulted from this technique.     

A technique for the quantification of Duddingtonia flagrans chlamydospores in sheep faeces

Previous observations showed that Duddingtonia flagrans chlamydospores were visualized in McMaster chambers containing faeces of treated sheep. This trial explored the McMaster technique as a tool to quantify chlamydospores in sheep faeces. A range of individual chlamydospore doses (from 19.5 x 10(6) to 177.5 x 10(6)) were offered orally to nine lambs for 7 consecutive days. A faecal sample (5 g) was daily obtained from the rectum of each animal (from days 1 to 13) to perform the McMaster technique using a sugar flotation fluid with 1.27 g/mL density. Each chlamydospore counted in the McMaster chamber was considered as 50 chlamydospores per g of faeces (CPG). The results confirmed that the estimated CPG was associated with the daily dose offered to the animals (r(2)=0.90; P<0.001). Furthermore, the total chlamydospore dose received by each animal was strongly associated to the total quantity of CPG obtained from the bulk faeces (TCtot) (r(2)=0.96; P<0.0001). Quantification of CPG can be used as a helpful tool to determine the number of chlamydospores reaching the faeces in orally dosed animals. This could be used to evaluate the efficacy of D. flagrans for the control of gastrointestinal nematode larvae in sheep faeces.     

Clinical assessment of gallinaceous birds and waterfowl in backyard flocks.

Obtaining a complete history and performing a physical examination are key steps in any clinical assessment. In addition, being familiar with common clinical signs helps in the formulation of a differential diagnosis. Once a definitive diagnosis is made, it is important to follow up each case with client education as well as the development of preventive medicine programs for avian patients.     

瘤胃总甲烷菌实时荧光定量PCR方法的构建

试验构建了瘤胃总甲烷菌实时荧光绝对定量PCR的标准品及标准曲线用于总甲烷菌的定量测定,并提取瘤胃微生物总DNA, 以甲烷菌特异性引物进行PCR扩增, 回收纯化PCR产物,与pBS-T载体连接并转化到大肠杆菌,再用氨苄西林培养基筛选阳性重组质粒, 提取含目的片段质粒DNA, 通过PCR及测序鉴定重组质粒.根据OD值确定浓度, 将梯度稀释的质粒作为模板, 进行荧光定量PCR反应并作出标准曲线.结果表明: 所构建的标准曲线具有很高的相关性(R2=0.992), 并获得了高扩增效率产物.     

Use of a constant-virus diluting-serum microneutralization technique for the detection and quantification of infectious bronchitis virus antibodies

A constant-virus diluting-serum microneutralization test (CVMNT) for avian infectious bronchits virus (IBV) was evaluated for both reliability and repeatability. The virus used in the assay was a chick kidney (CK) cell-adapted strain, the Beaudette strain (IBV 42). Sera tested were from 24-week-old broiler-breeder chickens that had been vaccinated 3 times from a combination vaccine of Newcastle disease virus (NDV) and IBV. Test results were not repeatable or comparable when the same sera were tested on different days, but test results were repeatable and comparable when the sera were tested on the same day. Differences in virus titer at the different times that tests were performed appeared to cause the variation in test results. A comparison was made between the CVMNT and a constant-serum diluting-virus microneutralization test (CSMNT). The CVMNT was able to detect differences in flock antibody titers that the CSMNT could not.     

微量元素添加剂防结块处理技术的试验研究

<正>微量元素添加剂经过粉碎处理后,密封贮存,以供生产预混合饲料之用。但是作为微量元素添加剂的硫酸铜、硫酸亚铁等硫酸盐经粉碎后,无论是直接贮存,还是与稀释剂混合后贮存,都出现了不同程度的结块现象(见表1、表2)。     

Development of a technique for serial bilateral renal biopsy in steers

A biopsy procedure was developed to provide serial kidney samples from standing steers. Ten clinically normal steers were given intramuscular injections of gentamicin sulfate, 4 mg/kg body weight. Renal biopsy was performed at 5 separate times. After feed was withheld for 24 h, laparoscopic surgery was performed in standing stocks. Acepromazine, xylazine, and butorphanol were used for sedation and analgesia, and 2% lidocaine was used for local anesthesia. Two incisions approximately 2 cm long were made in the paralumbar fossa to allow for trocar introduction. The abdomen was insufflated with CO2 and, with endoscopic guidance, a biopsy forceps used to remove a kidney sample 2 to 3 mm in diameter, by either a left or a right abdominal approach. Each operation was recorded on videotape, and images were also captured with a digital medical device system. Respiration, heart rate, temperature, appetite, attitude, and postural positions were evaluated at 12, 24, 48, and 72 h after surgery. The 51 laparoscopic procedures provided 48 renal samples (approximately 100 mg each). The 1st and 2nd samples were from the right kidney, and the 3rd sample was from either the left or the right kidney; the 4th and 5th samples were from the left kidney. Adhesions made an approach from the right side difficult for the 3rd sample. No clinical changes were observed in 9 steers after the procedure. One steer died after the 3rd procedure owing to hemorrhage.     

Clinical assessment and outcome of a single-layer technique for anastomosis of the small intestine in horses

In order to assess postoperative outcome in horses undergoing end-to-end anastomosis of the small intestine, performed using a one-layer technique, 15 horses that underwent exploratory coeliotomy, resection of the small intestine and end-to-end anastomosis using a continuous Lembert pattern were studied. Information on the age, breed, sex, diagnosis, treatment, complications and outcome of each case were obtained from medical records. Follow-up information was obtained via telephone conversations with clients and trainers. Five of the horses had short-term postoperative complications: one had postoperative ileus (POI), colic and peritonitis, one had POI and colic, two had POI only and one had diarrhoea only. A second exploratory coeliotomy was recommended in two of the 15 horses (13 per cent). The short-term survival rate, defined as survival up to the time of discharge from the hospital, was 93.3 per cent (14 of 15 horses). The long-term survival rate, defined as survival for at least 12 months after the surgery, was 84.6 per cent (11 of 13 horses followed up).     

Development of a fecal sample collection strategy for extraction and quantification of fecal immunoglobulin A in dogs

OBJECTIVE: To develop a fecal sample collection strategy and quantification method for measurement of fecal IgA concentrations in dogs. SAMPLE POPULATION: Fecal samples from 23 healthy pet dogs of various breeds. PROCEDURES: Immunoglobulin A was extracted from fecal samples. An ELISA for the measurement of fecal IgA concentrations was established and analytically validated. Intraindividual variation of fecal IgA was determined by calculation of coefficients of variation. A sample collection strategy was developed on the basis of results of intraindividual variation of fecal IgA concentrations. A reference range for fecal IgA concentrations was determined. RESULTS: The method for extraction and quantification of fecal IgA was determined to be sufficiently sensitive, reproducible, accurate, and precise. On the basis of the intraindividual variability of our results, the determined fecal sample collection strategy required analysis of a total of 4 fecal samples/dog, with each fecal sample collected on 2 consecutive days with 28 days between sample collection periods (ie, days 1 and 2 followed by days 28 and 29). Reference range values for fecal IgA concentration were 0.22 to 3.24 mg/g of feces. CONCLUSIONS AND CLINICAL RELEVANCE: Methods of fecal IgA extraction and quantification used in our study allow for identification of dogs with consistently low fecal IgA concentrations. Use of these techniques will enable future investigations into possible associations between low fecal IgA concentrations and signs of gastrointestinal disease in dogs.     

Development of a retrobulbar injection technique for ocular surgery and analgesia in dogs

OBJECTIVE: To develop and compare 3 techniques for retrobulbar injection of local anesthetic agents for ocular surgery and analgesia in dogs. DESIGN: Prospective study. ANIMALS: 17 dogs (including 9 cadavers). PROCEDURES: Inferior-temporal palpebral (ITP), perimandibular, and combined superior-inferior peribulbar injection techniques were compared by assessing the distribution of latex after injection into the orbits of 5 canine cadavers; magnetic resonance imaging (MRI) evaluation of the distribution of contrast agent after injection in the retrobulbar space of 4 canine cadavers; and assessment of the efficacy and MRI evaluation of the anatomic distribution of injections of a lidocainecontrast agent mixture in 4 anesthetized, nonrecovery dogs. By use of the preferred technique (ITP), the ocular effects of lidocaine anesthesia were evaluated in 4 dogs; during a 2-week period after treatment, dogs underwent ophthalmic examination, Schirmer tear testing (STT), intraocular pressure (IOP) measurement, and Cochet-Bonnet esthesiometry. RESULTS: Of the 3 techniques, the ITP technique was the preferred method for retrobulbar administration of anesthetic agent in dogs because it was efficacious (pupil dilation and central rotation of the globe achieved in all eyes), easiest to perform, and provided thorough coverage of the intraconal retrobulbar space without complication. During the 2-week follow-up period, the ITP injection did not significantly affect STT, IOP, or Cochet-Bonnet esthesiometry values in dogs. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs, retrobulbar administration of anesthetic agents via the ITP technique is a potential alternative to systemic administration of neuromuscular blocking agents for ophthalmic surgery and provides the additional benefit of local ocular analgesia.     

Development and analytic validation of a radioimmunoassay for the quantification of canine calprotectin in serum and feces from dogs

OBJECTIVE: To develop and analytically validate a radioimmunoassay (RIA) for the quantification of canine calprotectin (cCP) in serum and fecal extracts of dogs. Sample Population-Serum samples (n = 50) and fecal samples (30) were obtained from healthy dogs of various breeds and ages. PROCEDURES: A competitive, liquid-phase, double-antibody RIA was developed and analytically validated by assessing analytic sensitivity, working range, linearity, accuracy, precision, and reproducibility. Reference intervals for serum and fecal cCP concentrations were determined. RESULTS: Sensitivity and upper limit of the working range were 29 and 12,774 microg/L for serum and 2.9 and 1,277.4 microg/g for fecal extracts, respectively. Observed-to-expected ratios for serial dilutions of 6 serum samples and 6 fecal extracts ranged from 95.3% to 138.2% and from 80.9% to 118.1%, respectively. Observed-to-expected ratios for spiking recovery for 6 serum samples and 6 fecal extracts ranged from 84.6% to 121.5% and from 80.3% to 132.1%, respectively. Coefficients of variation for intra-assay and interassay variability were < 3.9% and < 8.7% for 6 serum samples and < 8.5% and < 12.6% for 6 fecal extracts, respectively. Reference intervals were 92 to 1,121 microg of cCP/L for serum and < 2.9 to 137.5 microg of cCP/g for fecal extracts. CONCLUSIONS AND CLINICAL RELEVANCE: The RIA described here was analytically sensitive, linear, accurate, precise, and reproducible for the quantification of cCP in serum and fecal extracts. This assay should facilitate research into the clinical use of serum and fecal cCP measurements in dogs with inflammatory bowel disease.     

Development and validation of a method for simultaneous separation and quantification of 5 different sugars in canine urine.

The objective of this project was to develop and validate a method for concurrent separation and quantification of methylglucose, rhamnose, xylose, sucrose, and lactulose in canine urine by using high pressure anion exchange liquid chromatography and pulsed amperometric detection. The method was validated by evaluating dilutional parallelism, spiking recovery, intra-assay variability, and inter-assay variability. Observed to expected ratios for 3 urine samples, and all sugars, ranged from 77.6% to 106.9% for a 1:2 dilution, 85.2% to 121.4% for a 1:4 dilution, and 91.6% to 163.7% for a 1:8 dilution. Observed to expected ratios for spiking recovery of 3 urine samples, all sugars, and 5 different spiking solutions, ranged from 85.5% to 116.7 % (mean +/- SD, 100.5 +/- 6.0%). The intra-assay coefficients of variation were 1.6%, 3.4%, and 4.7% for methylglucose; 1.6%, 2.0%, and 3.6% for rhamnose; 2.7%, 1.4%, and 1.1% for xylose; 9.8%, 3.4%, and 4.0% for sucrose; and 3.2%, 3.3%, and 3.3% for lactulose. Inter-assay coefficients of variation were 3.2%, 5.7%, and 4.2% for methylglucose; 4.3%, 5.4%, and 6.4% for rhamnose; 3.3%, 5.0%, and 4.2% for xylose; 9.4%, 9.9%, and 9.4% for sucrose; and 6.1%, 4.9%, and 2.7% for lactulose. In conclusion, a method for simultaneous separation and quantification of 5 sugars in canine urine was established and found to be linear, accurate, precise, and reproducible. This method may prove useful in the simultaneous evaluation of gastric permeability, small intestinal permeability, and small intestinal mucosal function in dogs with gastrointestinal disorders.