Vaccination with Mycobacterium bovis BCG Strains Danish and Pasteur in White-tailed Deer (Odocoileus virginianus) Experimentally Challenged with Mycobacterium bovis

Wildlife reservoirs of Mycobacterium bovis represent serious obstacles to the eradication of tuberculosis in domestic livestock and the cause for many faltering bovine tuberculosis eradication programs. One approach in dealing with wildlife reservoirs of disease is to interrupt inter-species and intraspecies transmission through vaccination of deer or cattle. To evaluate the efficacy of BCG vaccination in white-tailed deer, 35 deer were assigned to one of three groups; one s.c. dose of 107 CFU of M. bovis BCG Pasteur (n = 12); 1 s.c. dose of 107 CFU of M. bovis BCG Danish (n = 11); or unvaccinated deer (n = 12). After vaccination, deer were inoculated intratonsilarly with virulent M. bovis. Lesion severity scores of the medial retropharyngeal lymph node, as well as all lymph nodes combined, were reduced in vaccinated deer compared to unvaccinated deer. BCG Danish vaccinated deer had no late stage granulomas characterized by coalescent caseonecrotic granulomas containing numerous acid-fast bacilli compared to BCG Pasteur vaccinated or unvaccinated deer where such lesions were present. Both BCG strains were isolated as late as 250 days after vaccination from deer that were vaccinated but not challenged. In white-tailed deer, BCG provides protection against challenge with virulent M. bovis. Issues related to vaccine persistence, safety and shedding remain to be further investigated.

Vaccination of White-Tailed Deer (Odocoileus virginianus) with Mycobacterium bovis Bacillus Calmette Guerin

Wildlife reservoirs of Mycobacterium bovis represent serious obstacles to the eradication of tuberculosis in domestic livestock. In Michigan, USA tuberculous white-tailed deer transmit M. bovis to cattle. One approach in dealing with this wildlife reservoir is to vaccinate deer in order to interrupt the cycle of deer to deer and deer to cattle transmission. Thirty-one white-tailed deer were assigned to one of three groups; 2 SC doses of 107 CFU of M. bovis BCG (n = 11); 1 SC dose of 107 CFU of M. bovis BCG (n = 10); or unvaccinated deer (n = 10). After vaccination, deer were inoculated intratonsilarly with 300 CFU of virulent M. bovis. Gross lesion severity scores of the medial retropharyngeal lymph node were significantly reduced in deer receiving 2 doses of BCG compared to unvaccinated deer. Vaccinated deer had fewer lymph node granulomas than unvaccinated deer, and most notably, fewer late stage granulomas characterized by coalescent caseonecrotic granulomas containing numerous acid-fast bacilli. BCG was isolated from 7/21 vaccinated deer as long as 249 days after vaccination. In one case BCG was transmitted from a vaccinated deer to an unvaccinated deer. In white-tailed deer BCG provides measurable protection against challenge with virulent M. bovis. However, persistence of vaccine within tissues as well as shedding of BCG from vaccinates remain areas for further investigation.

Evaluation of an In Vitro Blood-Based Assay to Detect Production of Interferon-γ By Mycobacterium Bovis–Infected White-Tailed Deer (Odocoileus Virginianus)

Tuberculosis due to Mycobacterium bovis in captive Cervidae was identified as an important disease in the United States in 1990 and prompted the addition of captive Cervidae to the USDA Uniform Methods and Rules for eradication of bovine tuberculosis. As well, M. bovis infection was identified in free-ranging white-tailed deer in northeast Michigan in 1995. Tuberculosis in both captive and free-ranging Cervidae represents a serious challenge to the eradication of M. bovis infection from the United States. Currently, the only approved antemortem tests for tuberculosis in Cervidae are the intradermal tuberculin skin test and the blood tuberculosis test (BTB). At present, the BTB is not available in North America. Tuberculin skin testing of Cervidae is time-consuming and involves repeated animal handling and risk of injury to animals and humans. This study evaluated the potential of a new blood-based assay for tuberculosis in Cervidae that would decrease animal handling, stress, and losses due to injury. In addition, a blood-based assay could provide a more rapid diagnosis. Twenty 6–9-month-old white-tailed deer, male and female, were experimentally inoculated by instillation of 300 colony-forming units of M. bovis in the tonsillar crypts. Seven, age-matched uninfected deer served as controls. Blood was collected on days 90, 126, 158, 180, 210, 238, 263, and 307 after inoculation and was analyzed for the production of interferon-γ (IFN-γ) in response to incubation with M. bovis purified protein derivative (PPDb), M. avium PPDa, pokeweed mitogen (PWM), or media alone. Production of IFN-g in response to PPDb was significantly greater (P < 0.05) at all time points in samples from M. bovis–infected deer as compared with uninfected control deer, whereas IFN-γ production to PWM did not differ significantly between infected and control deer. Measurement of IFN-γ production to PPDb may serve as a useful assay for the antemortem diagnosis of tuberculosis in Cervidae.

Bovine Tuberculosis in Michigan Wildlife and Livestock

Since 1994, the state of Michigan has recognized a problem with bovine tuberculosis (TB), caused by Mycobacterium bovis, in wild white-tailed deer from a 12-county area in northeastern Lower Michigan. A total of 65,000 free-ranging deer have been tested, and 340 have been found to be positive for M. bovis. The disease has been found in other wildlife species, and, in 1998, in domestic cattle, where to date 13 beef cattle and 2 dairy cattle herds have been diagnosed with bovine TB. Unfortunately, the situation is unique in that there have never been reports of self-sustaining bovine TB in a wild, free-ranging cervid population in North America. Scientists, biologists, epidemiologists, and veterinarians who have studied this situation have concluded that the most logical theory is that high deer densities and the focal concentration caused by baiting (the practice of hunting deer over feed) and feeding are the factors most likely responsible for the establishment of self-sustaining TB in free-ranging Michigan deer. Baiting and feeding have been banned since 1998 in counties where the disease has been found. In addition, the deer herd has been reduced by 50% in the endemic area with the use of unlimited antlerless permits. The measures of apparent TB prevalence have been decreased by half since 1997, providing hopeful preliminary evidence that eradication strategies are succeeding.

Biology and Impacts of Pacific Island Invasive Species. 5. Eleutherodactylus coqui, the Coqui Frog (Anura: Leptodactylidae)

The nocturnal, terrestrial frog Eleutherodactylus coqui, known as the Coqui, is endemic to Puerto Rico and was accidentally introduced to Hawai‘i via nursery plants in the late 1980s. Over the past two decades E. coqui has spread to the four main Hawaiian Islands, and a major campaign was launched to eliminate and control it. One of the primary reasons this frog has received attention is its loud mating call (85–90 dB at 0.5 m). Many homeowners do not want the frogs on their property, and their presence has influenced housing prices. In addition, E. coqui has indirectly impacted the floriculture industry because customers are reticent to purchase products potentially infested with frogs. Eleutherodactylus coqui attains extremely high densities in Hawai‘i, up to 91,000 frogs ha-1, and can reproduce year-round, once every 1–2 months, and become reproductive around 8–9 months. Although the Coqui has been hypothesized to potentially compete with native insectivores, the most obvious potential ecological impact of the invasion is predation on invertebrate populations and disruption of associated ecosystem processes. Multiple forms of control have been attempted in Hawai‘i with varying success. The most successful control available at this time is citric acid. Currently, the frog is established throughout the island of Hawai‘i but may soon be eliminated on the other Hawaiian Islands via control efforts. Eradication is deemed no longer possible on the island of Hawai‘i.

Landscape-Genetic Analysis of Population Structure in the Texas Gray Fox Oral Rabies Vaccination Zone

In west-central Texas, USA, abatement efforts for the gray fox (Urocyon cinereoargenteus) rabies epizootic illustrate the difficulties inherent in large-scale management of wildlife disease. The rabies epizootic has been managed through a cooperative oral rabies vaccination program (ORV) since 1996. Millions of edible baits containing a rabies vaccine have been distributed annually in a 16-km to 24-km zone around the perimeter of the epizootic, which encompasses a geographic area >4 x 105 km2. The ORV program successfully halted expansion of the epizootic into metropolitan areas but has not achieved the ultimate goal of eradication. Rabies activity in gray fox continues to occur periodically outside the ORV zone, preventing ORV zone contraction and dissipation of the epizootic. We employed a landscape-genetic approach to assess gray fox population structure and dispersal in the affected area, with the aim of assisting rabies management efforts. No unique genetic clusters or population boundaries were detected. Instead, foxes were weakly structured over the entire region in an isolation by distance pattern. Local subpopulations appeared to be genetically non-independent over distances >30 km, implying that long-distance movements or dispersal may have been common in the region. We concluded that gray foxes in west-central Texas have a high potential for long-distance rabies virus trafficking. Thus, a 16-km to 24-km ORV zone may be too narrow to contain the fox rabies epizootic. Continued expansion of the ORV zone, although costly, may be critical to the long-term goal of eliminating the Texas fox rabies virus variant from the United States.

Eradicating Rats on Lehua Island, Hawaii, with the Help of GIS and GPS

Highlights • Wildlife Services used GIS and GPS to document and track bait distribution during each bait drop. • GIS and GPS were critical in making this eradication project effective and environmentally safe. • Use of the technologies ensured the coverage necessary for the project's goals.

Potential Attractants for Detecting and Removing Invading Gambian Giant Pouched Rats (Cricetomys gambianus)

BACKGROUND: Native to Africa, Gambian giant pouched rats (Gambian rats; Cricetomys gambianus Waterh.) are a threatening invasive species on a Florida island, Grassy Key. Gambian giant pouched rats shifted from a domestic pet to invading species after suspected release from a pet breeder. Because of the large size of Gambian rats (weighing up to 2.8 kg), they pose a serious threat to native species (particularly nesting species) and agricultural crops, especially if Gambian rats invade mainland Florida. Also, Gambian rats pose a threat from disease, as they were implicated in a monkeypox outbreak in the mid-western United States in 2003. The United States Department of Agriculture’s Wildlife Services has initiated eradication and detection efforts in the Florida Keys, but trapping the sparse population of Gambian rats has proven difficult. RESULTS: Fifteen attractants that could be used in traps for capturing or detecting single or paired Gambian rats were tested. It was found that conspecific scents (i.e. feces and urine) from other Gambian rats were the best treatment for attracting single and paired Gambian rats. Single Gambian rats explored more attractant types than paired Gambian rats. CONCLUSIONS: Effective attractants for use with Gambian rats have been identified, and multiple attractant types should be used to capture or detect the sparse population. It is recommended that mainly urine and feces from Gambian rats be used, but peanut butter, anise, ginger and fatty acid scent could also be useful for attracting the currently small population of Gambian rats on Grassy Key.

Taking an Endemically Infected Bovine Tb Area Through to Livestock Clearance. A Case Study of West Taupo, New Zealand

The Pest Management Strategy for Bovine Tuberculosis (Tb) in New Zealand aims to achieve efficient freedom from Tb by 2013 and to eradicate the disease from livestock and wildlife. The West Taupo area, in the central North Island of New Zealand, was chronically infected with Tb in both domestic livestock herds (cattle and deer) and within wildlife populations (brushtail possum, ferret, feral deer and pigs). Through the development and implementation of a technically innovative management plan, this area is now approaching Tb free status. The case study / management plan reported here discusses the operational techniques and strategies that were implemented to achieve Tb clearance in the livestock herds and the possibilities of eradication from wildlife species. It particularly identifies the variations in control strategies that are required as population densities reduce and the challenges of maintaining strong effective control at low densities of some wildlife species, whilst not needing to control other species that were initially clinically diagnosed with Tb control. Use of diagnostic tools and education as an area moves through the cycle towards Tb freedom are as essential as the physical control activities. The use of intensive monitoring of both livestock and wildlife species as trend and performance indicators and the need to educate farmers, hunters and other land use groups become increasingly important.

THE ORGANISATION OF RABBIT CONTROL (Oryctolagus cuniculus) IN WESTERN AUSTRALIA

ABSTRACT: Under Western Australian legislation, landholders have an obligation to control rabbits on their properties; local authorities the responsibility to supervise their work whilst the Agriculture Protection Board has a Statewide supervisory and co-ordination role. Prior to 1950 (when the Agriculture Protection Board was formed) the central role was in the hands of a Government department which, through lack of staff and money was unable to provide adequate supervision, and rabbits were in plague proportions. Since 1950, the Board has actively engaged in a vigorous policy aimed at tighter control and supervision. To enable this, the Board has entered into a voluntary scheme with local authorities whereby the role of local supervision of landholders is passed to staff employed by the Board, but jointly financed by the local authority and the Board. A contract poisoning service is also pro¬vided by the Agriculture Protection Board to any landholder who is unable or unwilling, to meet his obligations in this area. Both services are subsidised. Two of the major reasons for the poor level of control existing before 1950, have thereby been minimised. Soon after its formation, the Board set up a research section which has devoted nearly all of its activities to applied research on control of the State's many vertebrate pest problems. In the rabbit control area, poisoning has received most attention. The "One-Shot" method of poisoning was developed after years of research. Fumigation is at present being closely studied as is the economics of complete eradication from some areas of the State. Greatest needs in the applied rabbit research field at present are: (1) a selective poison, or poisoning regime, which will not harm stock, and (2) a more complete understanding of the economics of control and eradication. The serious rabbit problem which existed in 1950 has been reduced to very small proportions, by organisational development using local research findings. These organisational developments have been implemented by circumvention rather than confrontation.

PHENOLOGY AND FRUIT TRAITS OF ARCHONTOPHOENIX CUNNINGHAMIANA, AN INVASIVE PALM TREE IN THE ATLANTIC FOREST OF BRAZIL

The Australian palm Archontophoenix cunninghamiana was introduced into Brazil as an ornamental species, and became a dangerous invader of remnant Atlantic forest patches, demanding urgent management actions that require careful planning. Its fruits are greatly appreciated by generalist birds and its sudden eradication could be as harmful as its permanence in the native community. Our hypothesis was that A. cunninghamiana phenology and fruit traits would have facilitated the invasion process. Hence the aim of the study was to characterize the reproductive phenology of the palm by registering flowering and fruiting events, estimating fruit production, and evaluating fruit nutritional levels. Phenological observations were carried out over 12 months and analyzed statistically. Fruit traits and production were estimated. Pulp nutritional levels were determined by analyzing proteins, lipids, and carbohydrates. Results showed constant flowering and fruiting throughout the year with a weak reproductive seasonality. On average, 3651 fruits were produced per bunch mainly in the summer. Fruit analysis revealed low nutrient contents, especially of proteins and lipids compared with other Brazilian native palm species. We concluded that the abundant fruit production all year round, and fruit attractivity mainly due to size and color, :may act positively on the reproductive performance and effective dispersion of A. cunninghamiana. As a management procedure which would add quality to frugivore food resources we suggest the replacement of A. cunninghamiana by the native palm Euterpe edulis, especially in gardens and parks near to Atlantic forest fragments.

Antimicrobial activity and enterococcus faecalis biofilm formation on chlorhexidine varnishes

Objective: To evaluate, in vitro, the antimicrobial activity and biofilm formation of three chlorhexidine varnishes in four Enterococcus faecalis strains: E. faecalis ATCC 29212, E. faecalis EF-D1 (from failed endodontic treatment), E. faecalis 072 (cheese) and E. faecalis U-1765 (nosocomial infection), and one Enterococcus durans strain (failed endodontic treatment). Study Design: The direct contact test was used to study the antimicrobial activity. Bacterial suspensions were exposed for one hour to EC40, Cervitec (CE) and Cervitec Plus (CEP) varnishes. "Eradication" was defined as 100% bacterial kill. The formation of enterococci biofilms was tested on the surface of the varnishes after 24 hours of incubation and expressed as percentage of biofilm reduction. Results: EC40 eradicated all strains except E. faecalis ATCC 29212, where 98.78% kill was achieved. CE and CEP showed antimicrobial activity against all the strains, but most clearly against E. durans and E. faecalis 072. EC40 completely inhibited the formation of biofilm of E. faecalis ATCC 29212, E. faecalis 072 and E. durans. CE and CEP led to over 92% of biofilm reduction, except in the case of E. faecalis U-1765 on CEP (76.42%). Conclusion: The three varnishes studied were seen to be effective in killing the tested strains of enterococci and in inhibiting the formation of biofilm, the best results being observed with EC40.

Governança ambiental e economia verde

A Conferência Rio+20 mobiliza a comunidade global em 2012 para participar de um desafiador debate sobre a realidade ambiental global e modus operandi existente quanto à temática ampla e genérica do desenvolvimento e do ambiente. Um dos temas estruturantes desta reunião é a transição para uma economia verde no contexto do desenvolvimento sustentável e da erradicação da pobreza. O tema da Governança Ambiental Global um dos carros chefe do debate na Rio+20, no intuito de promover e acelerar a transição rumo a sociedades sustentáveis, configura a construção muitas vezes, de forma controversa, das condições para a definição de novos espaços institucionais e processos decisórios compartilhados. Este artigo propõe aos leitores uma reflexão para discutir que tipo de sustentabilidade está por trás da economia verde, a sua aplicabilidade e o que deva ser priorizada na discussão de governança ambiental. Isto se explica na medida em que existe a necessidade de mudar os mecanismos de utilização dos recursos, profundamente injustos, e que impedem avanços nos processos decisórios, pois as decisões de poucos tem configurado uma lógica perversa de expropriação de recursos naturais e não resolução da exclusão social.

Therapeutic DNA Vaccine Encoding Peptide P10 against Experimental Paracoccidioidomycosis

Paracoccidioidomycosis (PCM), caused by Paracoccidioides brasiliensis, is the most prevalent invasive fungal disease in South America. Systemic mycoses are the 10th most common cause of death among infectious diseases in Brazil and PCM is responsible for more than 50% of deaths due to fungal infections. PCM is typically treated with sulfonamides, amphotericin B or azoles, although complete eradication of the fungus may not occur and relapsing disease is frequently reported. A 15-mer peptide from the major diagnostic antigen gp43, named P10, can induce a strong T-CD4+ helper-1 immune response in mice. The TEPITOPE algorithm and experimental data have confirmed that most HLA-DR molecules can present P10, which suggests that P10 is a candidate antigen for a PCM vaccine. In the current work, the therapeutic efficacy of plasmid immunization with P10 and/or IL-12 inserts was tested in murine models of PCM. When given prior to or after infection with P. brasiliensis virulent Pb 18 isolate, plasmid-vaccination with P10 and/or IL-12 inserts successfully reduced the fungal burden in lungs of infected mice. In fact, intramuscular administration of a combination of plasmids expressing P10 and IL-12 given weekly for one month, followed by single injections every month for 3 months restored normal lung architecture and eradicated the fungus in mice that were infected one month prior to treatment. The data indicate that immunization with these plasmids is a powerful procedure for prevention and treatment of experimental PCM, with the perspective of being also effective in human patients.

Plant invasions research in Latin America: fast track to a more focused agenda

While many developed countries have invested heavily in research on plant invasions over the last 50 years, the immense region of Latin America has made little progress. Recognising this, a group of scientists working on plant invasions in Latin America met in Chile in late 2010 to develop a research agenda for the region based on lessons learned elsewhere. Our three main findings are as follows. (1) Globalisation is inevitable, but the resultant plant introductions can be slowed or prevented by effective quarantine and early intervention. Development of spatially explicit inventories, research on the invasion process and weed risk assessments can help prioritise and streamline action. (2) Eradication has limited application for plants and control is expensive and requires strict prioritisation and careful planning and evaluation. (3) Accepting the concept of novel ecosystems, new combinations of native and introduced species that no longer depend on human intervention, may help optimise invasive species management. Our vision of novel ecosystem management is through actions that: (a) maintain as much native biodiversity and ecosystem functionality as possible, (b) minimise management intervention to invasives with known impact, and (c) maximise the area of intervention. We propose the creation of a Latin American Invasive Plants Network to help focus the new research agenda for member countries. The network would coordinate research and training and establish funding priorities, develop and strengthen tools to share knowledge, and raise awareness at the community, governmental and intergovernmental levels about the social, economic and environmental costs of plant invasions.