Development and trial of a bovine herpesvirus 1 - thymidine kinase deletion virus as a vaccine

SUMMARY An Australian bovine herpesvirus 1 (BHV1) isolate with a defined (427 base pair) deletion in the protein coding region of the thymidine kinase gene was obtained by standard marker rescue procedures. After selection in the presence of the nucleotide analogue 5iodo-deoxy-uridine the virus was analysed by hybridisation with three differential oligonucleotide probes, restriction endonuclease profile studies and DNA sequence analysis. The virus elicited an immune response in recipient animals after either intramuscular or intravenous administration and produced no significant deleterious side-effects when administered at a dose sufficient to stimulate the host immune response. The safety and immunogenicity of the recombinant BHV1 virus 39B1 were similar to those reported for other registered BHV1 vaccines and the virus would appear to be suitable for the production of a vaccine seed lot and more exhaustive field trials as a prelude to commercial vaccine production and registration.     

Practices and opinions of New Zealand beef cattle farmers towards bovine viral diarrhoea control in relation to real and perceived herd serological status

ABSTRACT

Aims: To investigate the seroprevalence of infection with bovine viral diarrhoea (BVD) virus among 75 beef herds and seroconversion in cattle during early pregnancy, and to determine the practices and opinions of farmers towards BVD control and their association with real and perceived herd serological status.

Methods: Blood samples were collected before mating in 75 beef herds across New Zealand from 15 unvaccinated heifers that had delivered their first calf that season. Serum samples were tested for BVD antibodies using ELISA individually, and after pooling samples for each farm. Animals that were antibody-negative were retested at either pregnancy diagnosis or weaning. Farmers were asked to complete a detailed survey about herd demographics, BVD testing and vaccination practices, and opinions towards national BVD control.

Results: Based on the pooled serum antibody ELISA results, there were 28/75 (37%) negative herds, 15/75 (20%) suspect herds, and 32/75 (43%) positive herds. Of 1,117 animals sampled 729 (65.3%) tested negative for BVD virus antibodies; when retested, 47/589 (8.0%) animals from 13/55 (24%) herds had seroconverted. Among 71 famers providing survey responses 11 (15%) believed their herd was infected with BVD, 24 (34%) were unsure and 36 (51%) did not think their herd was infected. Only 19/71 (18%) farmers had performed any BVD testing within the past 5 years and 50/70 (71%) had not vaccinated any cattle for BVD. Support for national BVD eradication programme was strong in 51/71 (56%) respondents, but the biggest challenge to BVD control was considered to be famer compliance. Compared to farmers who did not think their herd was infected, more farmers who thought BVD was present in their herds had previously tested for BVD, would consider testing all replacement calves, and would support establishing a national BVD database; fewer would consider purchasing BVD tested or vaccinated cattle only.

Conclusions and clinical relevance: Only 15% of the beef farmers in this study believed their herds were infected with BVD virus and few of them had undertaken BVD screening. Nevertheless many were supportive of implementing a national BVD control programme. It is likely that the lack of farmer awareness around BVD and the failure of farmers to recognise the potential impacts in their herds are hindering progress in controlling the disease in New Zealand. There are opportunities for New Zealand veterinarians to be more proactive in helping beef farmers explore BVD management options.     

Reference range and repeatability of a combined intestinal permeability and function test in clinically healthy Irish setter dogs

A reference range was determined for a combined test of differential sugar permeability, using lactulose (L) and rhamnose (R), and intestinal function, using D-xlylose (X) and 3-O-methylglucose (G), in clinically healthy adult Irish setter dogs. Urinary L/R ratios in 48 Irish setters from one to 12 years old varied from 0·03 to 0·18, with a mean ( ) of 0·10 (0·01); X/G ratios varied from 0·46 to 0·81, with a mean ( ) of 0·59 (0·01). There were no significant differences between L/R or X/G ratios of dogs of different sex (P>0·2) or age (P0·5), using analysis of covariance. Lactulose/rhamnose ratios of ≥0·18 and X/G ratios ≥0·43 were considered normal, defined by the respective mean ± 2 . Repeatability was established by performing three permeability and function tests at monthly intervals in twelve of the dogs. Analysis of repeated L/R and X/G ratios by means of linear models procedure revealed no significant differences between measurements made on successive occasions (P>0·15), confirming the repeatability of this test.     

Identification of equine herpesvirus 3 (equine coital exanthema virus), equine gammaherpesviruses 2 and 5, equine adenoviruses 1 and 2, equine arteritis virus and equine rhinitis A virus by polymerase chain reaction

OBJECTIVE: To develop rapid (< 8 hour) tests using polymerase chain reaction (PCR) for the diagnosis of equine herpesvirus 3 (EHV3; equine coital exanthema virus), equine gammaherpesviruses 2 (EHV2) and EHV5, equine adenovirus 1 (EAdV1), EAdV2, equine arteritis virus (EAV), equine rhinitis A virus (ERAV; formerly equine rhinovirus 1) DESIGN: Either single round or second round (seminested) PCRs were developed and validated. METHODS: Oligonucleotide primers were designed that were specific for each virus, PCR conditions were defined and the specificity and sensitivity of the assays were determined. The application of the tests was validated using a number of independent virus isolates for most of the viruses studied. The PCRs were applied directly to clinical samples where samples were available. RESULTS: We developed a single round PCR for the diagnosis of EHV3, a seminested PCR for EHV2 and single round PCRs for EHV5, EAdV1, EAdV2 and RT-PCRs for EAV and ERAV. The PCR primer sets for each virus were designed and shown to be highly specific (did not amplify any recognised non-target template) and sensitive (detection of minimal amounts of virus) and, where multiple virus isolates were available all isolates were detected. CONCLUSION: The development and validation of a comprehensive panel of PCR diagnostic tests, predominantly for viruses causing equine respiratory disease, that can be completed within 8 hours from receipt of clinical samples, provides a major advance in the rapid diagnosis or exclusion diagnosis of these endemic equine virus diseases in Australia.     

提高辣椒离体再生不定芽伸长率的研究

通过调整培养基中的几种生长调节剂种类及其浓度组合，以及不同添加物，诱导不定芽分化与植株再生，有效地提高了不定芽的伸长率．结果表明：选用辣椒9～11d苗龄的带柄子叶在培养基MB＋BA5．0mg／L＋IAA1．0mg／L十GA31．0mg／L＋蔗糖3％＋琼脂6．5g／L十椰乳5％＋AgN035．0mg／L上培养，分化频率达97．8％；芽丛在MB＋ZT1．0mg／L＋IAA1．0mg／L＋GA31．5mg／L十椰乳5％＋AgN035．0mg／L培养基上伸长率达76．0％；幼苗在Ms＋IAA0．1mg／L＋NAA0．2mg／L培养基上能正常生根，生根率最高100％，并成长为健壮的再生植株。     

The absence of neuromas in beaks of adult hens after conservative trimming at hatch

Objective To determine the effects of the amount of beak removed and cauterisation time on neuroma formation in hens.
Design A pathology study with controls.
Animals Twenty domestic fowl were beak-trimmed. Three non-beak-trimmed domestic fowl were used as controls.
Procedure Beaks of two age groups with two levels of beak removal and either 2 s or 4 s cauterisation, were investigated macroscopically and microscopically for deformities.
Results Scattered trauma-associated neuromas were present in the beaks of pullets 10 weeks after moderate trimming at hatch. Neuromas were not present in beaks of adult hens that had been similarly trimmed. Sensory corpuscles were present 10 and 70 weeks after moderate trimming, though fewer in number than in intact control hens. In contrast, trauma-associated neuromas persisted in beaks of 70-week-old hens that had been severely trimmed at hatch. A range of deformities that were absent in moderately trimmed hens, were observed in hens with severely trimmed beaks. Receptors were not seen in severely trimmed beaks.
Conclusion Beak-trimming at hatch induces the formation of neuromas, regardless of the amount of tissue removed. There is a critical amount of beak tissue that can be removed, beyond which trauma-associated neuromas will not resolve, but will persist in mature hens.     

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.     

Diagnosis and management of malabsorption in dogs

Malabsorption can result from interference with either the degradation or absorption phases in the handling of dietary constituents and represents an important cause of weight loss and diarrhoea in dogs. Effective treatment depends on identification and understanding of the underlying disease which could affect the functional capacity of the exocrine pancreas or small intestine. Exocrine pancreatic insufficiency (EPI) can be identified by a low concentration of trypsin-like immunoreactivity in serum and results in serious malabsorption due to interference with degradation of carbohydrate, protein and fat. Treatment with oral pancreatic extract complemented by a low fat, high quality protein diet, is effective in many cases. Refractory cases may need additional treatment with an oral antibiotic for small intestinal bacterial overgrowth (SIBO), and H₂-receptor blockers to help prevent denaturation of the pancreatic extract by stomach acid. The pancreas plays a key role in the normal absorption of cobalamin (vitamin B₁₂) in dogs and malabsorption of cobalamin in EPI may not resolve with treatment so that cobalamin may need to be given parenterally. Small intestinal disease may result in interference with the number or functioning of individual enterocytes, in some cases accompanied by cellular infiltration of the mucosa. Diagnosis depends on indirect assessment of intestinal damage, for example by assay of serum vitamins and determination of intestinal absorption and permeability, and in selected cases followed by endoscopic examination, intestinal biopsy and culture of duodenal juice. Treatment depends on the disease and may include oral antibiotic for SIBO and immunosuppressive drugs for infiltrative disease. Dietary management is also important, for example with a restricted fat diet containing highly digestible carbohydrate and high quality protein, and when a dietary sensitivity is suspected a restriction diet of a selected protein source may be needed.     

低O 2和高CO 2对草莓采后酯类生物合成的作用模式

草莓(Fragria ananassa Duck.cv.Chandler)果实于采收当天分别放在:①空气;②0.25%O_2;③21%O_2+50%CO_2;④0.25%O_2+50%CO_2(用 N_2平衡)四种流动气体条件(5℃)下1～7d,以研究气调贮藏对草莓的挥发性物质和发酵性酶活性的影响。在三种气调贮藏条件下,果肉中乙醛、乙醇、乙酸乙酯、丁酸乙酯的浓度较空气处理明显增加而乙酸异丙酯和乙酸丁酯的含量下降。气调贮藏处理增加了丙酮酸脱羧酶(PDC)和乙醇脱氢酶(ADH)的活性但略微减少了醇酰基转移酶(即酯化酶,AAT)的活性。低 O_2、高 CO_2条件下草莓中乙醇的大量累积加速了乙酸乙酯的生物合成。由于 AAT 活性的降低及过量的乙醇与其他醇类化合物对相同羧基化合物的竞争作用导致其他乙酸酯类产物的减少。