Immunoproteomic analysis of the antibody response obtained in Nile tilapia following vaccination with a Streptococcus iniae vaccine

Streptococcus iniae is one of the most economically important Gram-positive pathogens in cultured fish species worldwide. The USDA-ARS Aquatic Animal Health Research Unit developed a modified (contains concentrated culture supernatant) S. iniae bacterin that has been demonstrated to be efficacious, and protection is mediated by specific anti-S. iniae antibodies. Although effective, the specific vaccine components important for efficacy are not known. In the present study, an immunoproteomic approach was utilized to identify whole-cell lysate proteins of S. iniae that stimulated specific antibody production in Nile tilapia (Oreochromis niloticus) following vaccination. Groups of tilapia were vaccinated by intraperitoneal injection with the modified S. iniae bacterin or were mock-vaccinated, and at 30 d post-vaccination sera samples were obtained from individual fish. Vaccination of tilapia with the S. iniae vaccine stimulated significantly elevated specific antibody responses against proteins of the bacterium and passive immunization of tilapia with this serum demonstrated the antibodies were highly protective. Whole-cell lysate proteins of S. iniae were separated by 2D-PAGE and were probed with a pooled serum sample from vaccinated tilapia. A total of eleven unique immunogenic proteins were positively identified by mass spectrometry. Based on research conducted on homologous proteins in other Streptococcus spp., antibodies specific for three of the identified proteins, enolase, glyceraldehyde-3-phosphate dehydrogenase, and fructose-bisphosphate aldolase, are likely involved in protection from streptococcosis caused by S. iniae.     

Identification and expression profile of multiple genes in Nile tilapia in response to formalin killed Streptococcus iniae vaccination

Twenty-eight expressed sequence tags (ESTs) were isolated from a Nile tilapia (Oreochromis niloticus) vaccinated vs non-vaccinated subtractive library at 12-h post injection of a formalin killed Streptococcus iniae ARS-98-60 vaccine. The 28 ESTs were classified in terms of their putative functions. Half of the ESTs identified were unknown proteins. Of the remaining half ESTs, 17% have putative functions in protein biosynthesis and 11% have putative functions in immunity, energy production, and signal transduction, respectively. Immunity-related ESTs identified included high density lipoprotein-binding protein vigilin, immunoglobulin heavy chain, and QM-like protein. Quantitative PCR revealed that one EST (cytochrome c oxidase subunit II) was highly upregulated (1825 ± 336 fold) in vaccinated fish compared to that in non-vaccinated fish. Of the remaining 27 ESTs, nine were significantly (P<0.05) upregulated (<20 fold) in vaccinated fish. The nine significantly upregulated genes included five unknown or hypothetical proteins and four known proteins (high density lipoprotein-binding protein vigilin, QM-like protein, ribosomal protein S13, and ribosomal protein L5). The upregulation of these genes induced by killed S. iniae vaccines suggest that they might play important role in Nile tilapia defense against S. iniae infection.     

Investigations towards an efficacious and safe strangles vaccine: submucosal vaccination with a live attenuated Streptococcus equi

As part of a search for a safe and efficacious strangles vaccine, several different vaccines and different vaccination routes were tested in foals. The degree of protection was evaluated after an intranasal challenge with virulent Streptococcus equi by clinical, postmortem and bacteriological examinations. Inactivated vaccines containing either native purified M-protein (500 microg per dose) or whole S equi cells (10(10) cells per dose) administered at least twice intramuscularly at intervals of four weeks, did not protect against challenge. Different live attenuated S equi mutants administered at least twice at intervals of four weeks by the intranasal route were either safe but not protective or caused strangles. In contrast, a live attenuated deletion mutant administered intramuscularly, induced complete protection but also induced unacceptable local reactions at the site of vaccination. Submucosal vaccination in the inner side of the upper lip with the live attenuated mutant at > or =10(8) colony-forming units per dose, appeared to be safe and efficacious in foals as young as four months of age. The submucosal vaccinations caused small transient swellings that resolved completely within two weeks, and postmortem no vaccine remnants or other abnormalities were found at the site of vaccination.     

Prevalence of intercurrent disease in dogs and cats presented for vaccination at a veterinary practice

A prospective study was undertaken to determine the prevalence of intercurrent disease in dogs and cats presented for vaccination in general practice. Fifty-two percent of animals were found to suffer from intercurrent disease and 3% had severe debilitating disease. A thorough clinical examination conducted at the time of routine vaccination appears to be an important element in maintaining companion animal health and welfare.     

Isolation of Streptococcus agalactiae and an aquatic birnavirus from doctor fish Garra rufa L

Background

The doctor fish, Garra rufa, has become increasingly popular as a treatment for skin disorders and for pedicures in recent years. Despite this there is very little information available regarding the welfare of these fish and the range of potential pathogens they may carry. In this study, a group of fish suffering from post-transport mortalities were examined and the isolated pathogens identified.

Findings

Group B Streptococcus agalactiae was isolated from kidney swabs of the fish and found to be resistant to a number of antibiotics. In addition to this, a fish virus belonging to the aquabirnavirus group, serogroup C was isolated for the first time in Ireland. However, no clinical signs of disease typical of bacterial or viral infections were observed in any fish examined.

Conclusions

As no clinical signs of disease attributable to either of the pathogens identified were found it was concluded that the mortalities were most likely due to transport related stress exacerbated by the presence of the pathogens. Further work is required to assess the suitability of current transport strategies and to examine the potential risk associated with the transport of live ornamental fish.     

Development of a multiplex PCR assay to detect Edwardsiella tarda,Streptococcus parauberis,and Streptococcus iniae in olive flounder (Paralichthys olivaceus)

A multiplex PCR protocol was established to simultaneously detect major bacterial pathogens in olive flounder (Paralichthys olivaceus) including Edwardsiella (E.) tarda, Streptococcus (S.) parauberis, and S. iniae. The PCR assay was able to detect 0.01 ng of E. tarda, 0.1 ng of S. parauberis, and 1 ng of S. iniae genomic DNA. Furthermore, this technique was found to have high specificity when tested with related bacterial species. This method represents a cheaper, faster, and reliable alternative for identifying major bacterial pathogens in olive flounder, the most important farmed fish in Korea.     

Policies for the vaccination of cats and dogs in New Zealand veterinary practices

AIMS: To determine current practices and attitudes towards vaccination of dogs and cats of veterinarians in New Zealand; the methods used for informing clients on which vaccines to use, and the preferred site for vaccination of cats.

METHODS: A postal questionnaire was sent to all 483 listed veterinary practices in New Zealand during February 2012. Some questions were specific to pet dogs, cats, or working farm dogs. Responses were categorised according to practice type and geographical region of the respondent. Factors associated with respondent recommendation of annual vaccination with modified live viral (MLV) vaccines were examined using logistic regression analysis. Vaccines that were considered to be essential for every animal were defined as core; those that may be recommended for animals whose location or lifestyle placed them at risk, were defined as non-core.

RESULTS: There were 204 useable returns, equivalent to a response rate of 42.2%, distributed across the country. Annual vaccination with MLV vaccines of dogs was recommended by 54/198 (27.3%) respondents, and of cats by 107/181 (59.1%) respondents. Factors associated with the recommendation of annual administration of MLV vaccines to dogs included being a companion animal practice, a desire for policies on vaccination to be left to individual clinics, and having one veterinarian in the practice. Administration of the final vaccination for puppies was recommended at ≥14 weeks old by 55/185 (29.7%) respondents, and for kittens at ≥13 weeks old by 42/183 (23%) respondents. Of respondents that administered MLV vaccines annually, 62/103 (60.2%) believed reducing the frequency of vaccination would reduce income, and 52/103 (50.5%) considered it would have a negative effect on animal health. Advice to enable clients to decide which non-core vaccines were administered was given by 181/199 (91%) respondents. Factors considered when recommending a vaccine included consideration of risk to individual patients (190/203; 93.6%), requirements of boarding kennels/catteries (165/203; 81.3%) and clinic vaccination policy (142/203; 70%). The preferred site for administering MLV vaccines to cats was the dorsal neck or inter-scapular region (137/198; 69.2%). Amongst respondents, 18 wanted disease surveillance information to allow for truly informed decisions to be made about vaccination.

CONCLUSIONS AND CLINICAL RELEVANCE: Veterinarians can now compare their own vaccination practices and attitudes with those of veterinarians nationally, and internationally. There is a need for national surveillance information and for continued education of the public and commercial kennel and cattery owners for optimal vaccination strategies to be developed.     

Development of a PCR assay for Streptococcus iniae based on the lactate oxidase (lctO) gene with potential diagnostic value

Streptococcus iniae is a well-known pathogen of both fish and humans that is difficult to identify by conventional biochemical tests. A PCR assay based on the lactate oxidase (lctO) gene of S. iniae was developed for the rapid and specific detection and identification of this pathogen from different sources. The PCR assay had a detection limit of 62-31 cells, and 25 pg of DNA per PCR reaction mixture. The PCR was also effective in detecting the bacterium from inoculated tissue homogenates, suggesting its potential use for a rapid and accurate diagnosis of S. iniae infections.     

Results of veterinary tumor research and their significance for practice]

Enzootic bovine leucosis and Marek's disease of fowl are the most important tumour diseases of farm animals, with the greatest repercussions on the national economy, today. Experimental findings obtained from either disease in recent years are reported in this paper, and an assessment is made of their importance to practice. In the context of enzootic bovine leucosis, particular reference is made to those findings which helped to establish the presence of virus infection, results by which the pathogen could be characterised, and to studies undertaken with the view to devising methods for virologico-serological diagnosis. In the context of Marek's disease, reference is made to virological, epizootiological, and diagnostic aspects, however, with emphasis being laid on possibilities for immunisation and, in that connection, on known fundamental concepts for successful immunoprophylaxis against that tumour disease.     

OLIVER: an online library of images for veterinary education and research

As part of a strategic move by the University of Sydney toward increased flexibility in learning, the Faculty of Veterinary Science undertook a number of developments involving Web-based teaching and assessment. OLIVER underpins them by providing a rich, durable repository for learning objects. To integrate Web-based learning, case studies, and didactic presentations for veterinary and animal science students, we established an online library of images and other learning objects for use by academics in the Faculties of Veterinary Science and Agriculture. The objectives of OLIVER were to maximize the use of the faculty's teaching resources by providing a stable archiving facility for graphic images and other multimedia learning objects that allows flexible and precise searching, integrating indexing standards, thesauri, pull-down lists of preferred terms, and linking of objects within cases. OLIVER offers a portable and expandable Web-based shell that facilitates ongoing storage of learning objects in a range of media. Learning objects can be downloaded in common, standardized formats so that they can be easily imported for use in a range of applications, including Microsoft PowerPoint, WebCT, and Microsoft Word. OLIVER now contains more than 9,000 images relating to many facets of veterinary science; these are annotated and supported by search engines that allow rapid access to both images and relevant information. The Web site is easily updated and adapted as required.     

Providing stewardship and leadership for the future of veterinary medical research: a responsibility of the veterinary colleges

As we contemplate responsibilities as well as opportunities in research, it is fair to begin by considering why research is, or should be, important to veterinary schools and colleges, to our profession, and to society. Veterinary research is conducted in many venues, such as colleges of veterinary medicine, veterinary science departments, comparative medicine departments, medical schools, and many other university departments, as well as in industry laboratories, governmental agencies, and other organizations. But schools and colleges of veterinary medicine have a unique and historic responsibility to ensure that our veterinary medical research programs are sufficient in quality, capacity, and depth to meet the research needs of our society. Consequently, my comments will be oriented toward veterinary schools and colleges.     

The experience of dog bites: a survey of veterinary science and veterinary nursing students

AIM: To investigate the circumstances of dogs biting people in New Zealand. METHODS: Veterinary science and veterinary nursing students enrolled at Massey University in 2003 completed a questionnaire detailing their lifetime experience of dog bites and their opinions of dog control legislation. RESULTS: The response rate was 100%. There were 228 completed responses to the survey, and 87 (38%) respondents had been bitten by a dog. Most bites caused minor injury, but 17 (20%) required medical attention. Male respondents were more likely to have been bitten. The highest proportion of respondents (27; 31%) had been bitten while between 6 and 10 years of age, and the hand was the part of the body most often injured. Eighteen (21%) bites involved a stranger's dog, but most bites were by a dog known to the victim. Respondents from rural areas were three times more likely to have been bitten than city dwellers. The victim's home was the place where most (31; 36%) bites occurred. Male dogs were responsible for a larger proportion of bites (38; 44%) than females (24; 28%); respondents were unsure about the sex of the remainder of the dogs (25; 29%) that bit them. Protecting the home, play, accidents, rough handling and pain were identified as the common reasons for dogs to bite. Most (46; 52%) respondents did not believe legislation could have prevented their bite. Only four (5%) bites were reported to authorities. CONCLUSION: Rural dwellers, people with more experience of dogs, children <11 years of age, and male respondents were bitten by dogs most often. Only legislation relating to the fencing of dog owners' properties may have prevented a reasonable proportion (25; 29%) of the bites reported here.     

Animal abuse and intimate partner violence: researching the link and its significance in Ireland - a veterinary perspective

Research on domestic violence has established a substantial association between intimate partner abuse and the abuse of children within the home. It is only recently however, that researchers have demonstrated the correlation between non-accidental injury in animals, and abuse of women by their intimate male partners. A growing body of evidence suggests that animal abuse can be an early indicator for other forms of violent behaviour. This research includes the responses of a sample of 23 women using refuge services in the Republic of Ireland. It investigates the connection between domestic violence and animal abuse, and ascertains if there is sufficient support service for animals and people relevant to domestic abuse. In the survey population, 57% of women reported witnessing one or more forms of abuse, or threats of abuse, of their pets. Five of which were reported to have resulted in the death of the pet. Eighty seven per cent of women felt a facility to accommodate pets would have made their decision to leave the family home easier. Four women disclosed that lack of such a service and concern for the welfare of their companion animals caused them to remain in their abusive relationships for longer than they felt appropriate. Nine families placed pets in the care of family or friends, one woman is unaware of the fate of her pet, while the pets of six families remained with the abusive male after his partner entered a refuge. The majority of women felt unable to talk to anyone about their fears for their pets' welfare. Many felt that there is no service which can provide temporary accommodation for womens' pets while they are in refuge. The results obtained support those found elsewhere in larger studies in the USA and UK, and demonstrate an association of animal abuse in households where there is reported domestic violence.     

Hierarchical Bayesian model for prevalence inferences and determination of a country's status for an animal pathogen

Certification that a country, region or state is "free" from a pathogen or has a prevalence less than a threshold value has implications for trade in animals and animal products. We develop a Bayesian model for assessment of (i) the probability that a country is "free" of or has an animal pathogen, (ii) the proportion of infected herds in an infected country, and (iii) the within-herd prevalence in infected herds. The model uses test results from animals sampled in a two-stage cluster sample of herds within a country. Model parameters are estimated using modern Markov-chain Monte Carlo methods. We demonstrate our approach using published data from surveys of Newcastle disease and porcine reproductive and respiratory syndrome in Switzerland, and for three simulated data sets.     

Developing and fostering a dynamic program for training in veterinary pathology and clinical pathology: veterinary students to post-graduate education

Recent reports project a deficiency of veterinary pathologists, indicating a need to train highly qualified veterinary pathologists, particularly in academic veterinary medicine. The need to provide high-quality research training for veterinary pathologists has been recognized by the veterinary pathology training program of the Ohio State University (OSU) since its inception. The OSU program incorporates elements of both residency training and graduate education into a unified program. This review illustrates the components and structure of the training program and reflects on future challenges in training veterinary pathologists. Key elements of the OSU program include an experienced faculty, dedicated staff, and high-quality students who have a sense of common mission. The program is supported through cultural and infrastructure support. Financial compensation, limited research funding, and attractive work environments, including work-life balance, will undoubtedly continue to be forces in the marketplace for veterinary pathologists. To remain competitive and to expand the ability to train veterinary pathologists with research skills, programs must support strong faculty members, provide appropriate infrastructure support, and seek active partnerships with private industry to expand program opportunities. Shortages of trained faculty may be partially resolved by regional cooperation to share faculty expertise or through the use of communications technology to bridge distances between programs. To foster continued interest in academic careers, training programs will need to continue to evolve and respond to trainees' needs while maintaining strong allegiances to high-quality pathology training. Work-life balance, collegial environments that foster a culture of respect for veterinary pathology, and continued efforts to reach out to veterinary students to provide opportunities to learn about the diverse careers offered in veterinary pathology will pay long-term dividends for the future of the profession.     

Skills and competencies required by veterinary pharmacologists: a blueprint for graduate education in veterinary pharmacology in North America

This report describes the results of two efforts to gather data on the needs of veterinary pharmacology, whether academic or industrial, to better define the skills and competencies that graduate programs in veterinary pharmacology should endeavor to provide. Data collected from an on-line survey of the animal health industry were used to develop a workshop at the biennial meeting of the American Academy of Veterinary Pharmacology and Therapeutics. Lists of skills were developed and categorized, and differences among the skills identified by industry vs. academy were outlined. This report offers a blueprint for graduate education in veterinary pharmacology in terms of the types and details of skills and competencies to better prepare future veterinary pharmacologists.