The Potential for Transmission of BCG from Orally Vaccinated White-Tailed Deer (Odocoileus virginianus) to Cattle (Bos taurus) through a Contaminated Environment: Experimental Findings

White-tailed deer (Odocoileus virginianus) experimentally infected with a virulent strain of Mycobacterium bovis have been shown to transmit the bacterium to other deer and cattle (Bos taurus) by sharing of pen waste and feed. The risk of transmission of M. bovis bacille Calmette-Guerin (BCG) vaccine from orally vaccinated white-tailed deer to other deer and cattle, however, is not well understood. In order to evaluate this risk, we orally vaccinated 14 white-tailed deer with 1×10⁹ colony forming units BCG in lipid-formulated baits and housed them with nine non-vaccinated deer. Each day we exposed the same seven naïve cattle to pen space utilized by the deer to look for transmission between the two species. Before vaccination and every 60 days until the end of the study, we performed tuberculin skin testing on deer and cattle, as well as interferon-gamma testing in cattle, to detect cellular immune response to BCG exposure. At approximately 27 weeks all cattle and deer were euthanized and necropsied. None of the cattle converted on either caudal fold, comparative cervical tests, or interferon-gamma assay. None of the cattle were culture positive for BCG. Although there was immunological evidence that BCG transmission occurred from deer to deer, we were unable to detect immunological or microbiological evidence of transmission to cattle. This study suggests that the risk is likely to be low that BCG-vaccinated white-tailed deer would cause domestic cattle to react to the tuberculin skin test or interferon-gamma test through exposure to a BCG-contaminated environment.     

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

Wildlife reservoirs of Mycobacterium bovis represent serious obstacles to the eradication of tuberculosis from livestock, particularly cattle. In Michigan, USA tuberculous white-tailed deer transmit M. bovis to other deer and cattle. One approach in dealing with this wildlife reservoir is to vaccinate deer, thus interfering with the intraspecies and interspecies transmission cycles. Thirty-three white-tailed deer were assigned to one of two groups; oral vaccination with 1×10⁸ colony-forming units of M. bovis BCG Danish (n = 17); and non-vaccinated (n = 16). One hundred eleven days after vaccination deer were infected intratonsilarly with 300 colony-forming units of virulent M. bovis. At examination, 150 days after challenge, BCG vaccinated deer had fewer gross and microscopic lesions, fewer tissues from which M. bovis could be isolated, and fewer late stage granulomas with extensive liquefactive necrosis. Fewer lesions, especially those of a highly necrotic nature should decrease the potential for dissemination of M. bovis within the host and transmission to other susceptible hosts.     

Evaluation of attractant flavours for use in oral vaccine baits for badgers (<Emphasis Type="Italic">Meles meles</Emphasis>)

European badgers (Meles meles) are a wildlife reservoir for Mycobacterium bovis infection (tuberculosis) in Ireland and the UK and are implicated in the transmission of infection to livestock. Vaccination of badgers with the human BCG vaccine (Bacille Calmette Guerin) is considered as an important strategy to reduce the burden of disease in this species, and a pragmatic approach is likely to involve oral vaccination. In this study, we evaluated nine different flavours for use as attractants in a prototype oral vaccine bait for European badgers (M. meles): aniseed, apple, cocoa powder, carob powder, curry, fish, garlic, peanut and strawberry. The bait matrix was composed of a natural lipid formulation, developed as a vehicle for oral vaccination against tuberculosis in wildlife. A ‘food for work’ paradigm was employed during the trials to ensure the animals were actively seeking the baits. The trials showed carob and cocoa powders were equally attractive and more attractive than any of the other candidates. Carob and cocoa show potential as bait attractants for badgers and might form part of a novel vaccine delivery system.     

Efficacy of oral and parenteral routes of Mycobacterium bovis bacille Calmette-Guerin vaccination against experimental bovine tuberculosis in white-tailed deer (Odocoileus virginianus): a feasibility study

We investigated the efficacy of oral and parenteral Mycobacterium bovis bacille Calmette-Guerin Danish strain 1331 (BCG) in its ability to protect white-tailed deer (Odocoileus virginianus) against disease caused by M. bovis infection. Twenty-two white-tailed deer were divided into four groups. One group (n=5) received 10(9) colony-forming units (cfu) BCG via a lipid-formulated oral bait; one group (n=5) received 10(9) cfu BCG in culture directly to the oropharynx, one group (n=6) was vaccinated with 10(6) cfu BCG subcutaneously, and one group served as a control and received culture media directly to the oropharynx (n=6). All animals were challenged 3 mo after vaccination. Five months postchallenge the animals were examined for lesions. Results indicate that both oral forms of BCG and parenterally administered BCG offered significant protection against M. bovis challenge as compared to controls. This study suggests that oral BCG vaccination may be a feasible means of controlling bovine tuberculosis in wild white-tailed deer populations.     

Species-specific visitation and removal of baits for delivery of pharmaceuticals to feral swine

Within the domestic swine industry there is growing trepidation about the role feral swine (Sus scrofa) play in the maintenance and transmission of important swine diseases. Innovative disease management tools for feral swine are needed. We used field trials conducted in southern Texas from February to March 2006 to compare species-specific visitation and removal rates of fish-flavored and vegetable-flavored baits with and without commercially available raccoon (Procyon lotor) repellent (trial 1) and removal rates of baits deployed in a systematic and cluster arrangement (trial 2). During trial 1, 1) cumulative bait removal rates after four nights ranged from 93% to 98%; 2) bait removal rates by feral swine, raccoons, and collared peccaries (Pecari tajacu) did not differ by treatment; and 3) coyotes (Canis latrans) removed more fish-flavored baits without raccoon repellent and white-tailed deer removed more vegetable-flavored baits without raccoon repellent than expected. During trial 2, feral swine removed fish-flavored baits distributed in a cluster arrangement (eight baits within 5 m2) at a rate greater than expected. Our observed bait removal rates illustrate bait attractiveness to feral swine. However, the diverse assemblage of omnivores in the United States compared with Australia where the baits were manufactured adds complexity to the development of a feral swine-specific baiting system for pharmaceutical delivery.     

Oral rabies vaccine (ORV) bait uptake by captive striped skunks

Aerial delivery of oral rabies vaccine (ORV) baits has proven effective in large-scale efforts to immunize wildlife against rabies, and in North America this strategy currently is being used to immunize foxes (Urocyon cinereoargenteus and Vulpes vulpes), raccoons (Procyon lotor), and coyotes (Canis latrans). Skunks are also a major reservoir and vector of rabies, but at present oral vaccines for use in skunks are not licensed. Furthermore, given differences in morphology (smaller jaws) and behavior (food handling and consumption), it is unknown if baits currently used in ORV campaigns would be effective for skunks. Because oral vaccine delivery is contingent upon puncture of the vaccine container (VC), baits need to be sufficiently attractive to elicit selection and consumption. Manipulation of the bait to facilitate vaccine ingestion by the target species is a critical element for an effective ORV bait. The objectives of this study were to assess manipulation and consumption of current ORV baits by striped skunks (Mephitis mephitis). We conducted four independent trials with penned animals and various baits to assess bait selection frequency, VC puncture frequency, and consumption. Video recorded trials were used to assess attractiveness of baits and consumption behavior of skunks. Bait characteristics, such as texture, size, and flavor influenced selection and consumption. Fish and chicken flavors were preferred and vaccine containers within selected baits were likely to be punctured. Vaccine ingestion seemed more likely if VCs were directly coated with the bait matrix. To make baits attractive to skunks and to ensure puncture of the VC, modifications to current baits should consider a smaller size, a meat-flavored matrix, a slightly pressurized VC, and a direct coating of matrix on the VC.     

A new flavor-coated sachet bait for delivering oral rabies vaccine to raccoons and coyotes

Research was conducted during 1996-2000 to develop baits for delivering an oral rabies vaccine to raccoons (Procyon lotor) and coyotes (Canis latrans). A bait was sought that: (1) was attractive to the target species, (2) could be distributed by aircraft, (3) was as effective (or more so) than the currently used fish meal polymer bait, and (4) could be produced in large numbers by automated procedures and could be purchased by user groups at substantially lower cost. Ten field trials were conducted to document raccoons' bait flavor preferences, evaluate a new vaccine sachet bait coated with various attractants, and determine if the sachet bait would effectively deliver Raboral V-RG oral rabies vaccine (Merial Limited, Athens, Georgia, USA) to this species. Raccoons preferred fish and crustacean-based flavors over those derived from plant materials. Raccoon visits to tracking stations, frequency of bait removals, and percent of sachets discarded by this species that were emptied of placebo vaccine indicated efficacy of the new bait was equal or superior to the currently used fish meal polymer bait. A field trial conducted in fall 1998 compared aerially distributed vaccine-laden sachet and polymer baits and showed there was no difference between the percent of raccoons from the test and reference areas subsequently found positive for rabies antibody. Four bait trials to determine coyote response to sachet baits were conducted in 1997-98. The propensity for canids to gulp or bolt smaller food items is well known. Thus, a first trial involved offering fish-flavored sachet baits of different sizes to 30 captive coyotes to determine if smaller size baits were more frequently swallowed intact. Two field trials were also conducted in fall 1997 to determine if free-ranging coyotes discriminated among sachet baits coated with different attractants. Finally, Raboral V-RG-laden poultry-flavored sachet baits were aerially dropped and the percent of seropositive coyotes was compared with coyotes from surrounding areas where fish meal polymer vaccine baits had been distributed. Captive coyotes did not swallow sachet baits intact, regardless of size. Bait preference field trials indicated that coyotes preferred poultry, cheese/beef tallow, and fish-flavored sachet baits and that such baits were taken at the same rate as polymer baits. A sample of coyotes from the area baited with vaccine-laden sachet baits had a markedly higher (P = 0.01) seropositivity rate than coyotes from areas where vaccine was distributed in polymer baits. Sachet bait production could be facilitated by automated technology and sachet baits used either as an alternative vaccine delivery device or in combination with the fish meal polymer bait.     

Detection of natural infection with Mycobacterium intracellulare in healthy wild-caught Chacma baboons (Papio ursinus) by ESAT-6 and CFP-10 IFN-γ ELISPOT tests following a tuberculosis outbreak

Background

The current tuberculosis vaccine is a live vaccine derived from Mycobacterium bovis and attenuated by serial in vitro passaging. All vaccine substrains in use stem from one source, strain Bacille Calmette-Guérin. However, they differ in regions of genomic deletions, antigen expression levels, immunogenicity, and protective efficacy.

Results

As a RecA phenotype increases genetic stability and may contribute restricting the ongoing evolution of the various BCG substrains while maintaining their protective efficacy, we aimed to inactivate recA by allelic replacement in BCG vaccine strains representing different phylogenetic lineages (Pasteur, Frappier, Denmark, Russia). Homologous gene replacement was achieved successfully in three out of four strains. However, only illegitimate recombination was observed in BCG substrain Russia. Sequence analyses of recA revealed that a single nucleotide insertion in the 5' part of recA led to a translational frameshift with an early stop codon making BCG Russia a natural recA mutant. At the protein level BCG Russia failed to express RecA.

Conclusion

According to phylogenetic analyses BCG Russia is an ancient vaccine strain most closely related to the parental M. bovis. We hypothesize that recA inactivation in BCG Russia occurred early and is in part responsible for its high degree of genomic stability, resulting in a substrain that has less genetic alterations than other vaccine substrains with respect to M. bovis AF2122/97 wild-type.     

Translating Bait Preference to Capture Success of Northern White-Tailed Deer

ABSTRACT Wildlife management and research have depended upon trapping as an essential tool for decades. Although deer (Odocoileus spp.) capture by Clover traps remains a basic technique that has changed little over time, researchers use it as an integral part of field operations to support increasingly sophisticated and costly project objectives. Despite reports of deer preference for certain baits, no study has determined if bait preference can effectively increase capture success of free-ranging deer. By supplementing corn bait with salt, peanut butter, or molasses, we tested effects of these bait treatments on capture success of free-ranging white-tailed deer (O. virginianus), as well as levels of nontarget animal disturbance in Clover traps, during February-March 2005. With 1,446 adjusted trap-nights and a 6.5% capture success rate, the probability of capture increased over time and varied among 4 study sites (df = 3, P < 0.005); however, we did not detect a significant effect of bait supplementation on capture success (df = 3, P > 0.8). Nontarget animal activity in the trap varied by site (df = 3, P < 0.001), bait treatment (df = 3, P=0.04), and Julian date (df=3, P < 0.001). Our results are the first to suggest that bait preference may not translate into actual improved capture success of free-ranging deer. Future research should focus on testing additional baits or bait supplements to determine if an increase in trapping success and a minimization of trap disturbance by nontarget species occurs.     

Specificity and success of oral-bait delivery to Eurasian wild boar in Mediterranean woodland habitats

Eurasian wild boar (Sus scrofa) is an important reservoir host for pathogens affecting humans and domestic animals. The eradication of these diseases may require the development of control strategies that reduce pathogen transmission between wildlife and domestic animals. Baiting for oral vaccine delivery is often considered for wildlife disease control. The effective and efficient field vaccination of wildlife requires species-specific baits as delivery vehicles for oral vaccines and designing appropriate baiting strategies. The objective of this study was to determine the proportion of young and adult wild boar and non-target animals that consumed baits containing a chemical marker, iophenoxic acid (IPA), in delivery trials conducted in summer in four different sites in the Mediterranean region of Spain where wild boar are abundant. The proportion of wild boar showing IPA markers in serum in autumn ranged from 11.5% to 56.4%. When attending to age classes, 12.6% to 72.7% of young individuals presented IPA. The results evidenced that the percent of wild boar that ingested the baits varied among study sites and age classes. Placing baits inside selective cages (for juveniles) and under heavy pavel stones (for adults) contributed to improve age specificity in bait consumption. We suggest ways for improving the age specificity of bait delivery systems used for young and adult wild boar.     

Fecal Volatile Organic Ccompound Profiles from White-Tailed Deer (Odocoileus virginianus) as Indicators of Mycobacterium bovis Exposure or Mycobacterium bovis Bacille Calmette-Guerin (BCG) Vaccination

White-tailed deer (Odocoileus virginianus) serve as a reservoir for bovine tuberculosis, caused by Mycobacterium bovis, and can be a source of infection in cattle. Vaccination with M. bovis Bacille Calmette Guerin (BCG) is being considered for management of bovine tuberculosis in deer. Presently, no method exists to non-invasively monitor the presence of bovine tuberculosis in deer. In this study, volatile organic compound profiles of BCG-vaccinated and non-vaccinated deer, before and after experimental challenge with M. bovis strain 95–1315, were generated using solid phase microextraction fiber head-space sampling over suspended fecal pellets with analysis by gas chromatography/mass spectrometry. Chromatograms were processed using XCMS Online to characterize ion variation among treatment groups. The principal component scores resulting from significant (α = 0.05) ion responses were used to build linear discriminant analysis models. The sensitivity and specificity of these models were used to evaluate the feasibility of using this analytical approach to distinguish within group comparisons between pre- and post-M. bovis challenge: non-vaccinated male or female deer, BCG-vaccinated male deer, and the mixed gender non-vaccinated deer data. Seventeen compounds were identified in this analysis. The peak areas for these compounds were used to build a linear discriminant classification model based on principal component analysis scores to evaluate the feasibility of discriminating between fecal samples from M. bovis challenged deer, irrespective of vaccination status. The model best representing the data had a sensitivity of 78.6% and a specificity of 91.4%. The fecal head-space sampling approach presented in this pilot study provides a non-invasive method to discriminate between M. bovis challenged deer and BCG-vaccinated deer. Additionally, the technique may prove invaluable for BCG efficacy studies with free-ranging deer as well as for use as a non-invasive monitoring system for the detection of tuberculosis in captive deer and other livestock.     

Isocitrate lyase encoding plasmids in BCG cause increased survival in ApoB100-only LDLR−/− mice

We studied the role of isocitrate lyase in the interaction between Mycobacterium bovis BCG and mice. ApoB100-only LDLR^?/? (B6;129S-ApoB^tm2SgyLdlr^tm1Her/J) mice were inoculated with M. bovis BCG harbouring plasmids carrying the gene for isocitrate lyase. The presence of ~29 times more copies of this gene resulted in a higher bacterial yield in the spleens and lungs of the infected mice. The spleen was 3–4 times heavier, and in the spleen the bacteria survived over 10 days longer than did the bacteria with the control plasmid. Propionate was less toxic for bacteria carrying icl plasmids in vitro. This recombinant BCG can be a possible vaccine candidate.     

Evaluating and predicting risk to a large reptile (Varanus varius) from feral cat baiting protocols

Control of introduced predators to mitigate biodiversity impacts is a pressing conservation challenge. Across Australia feral cats (Felis catus) are a major threat to terrestrial biodiversity. Currently feral cat control is hindered by the limited utility of existing predator baiting methods. Further proposed control methods include use of the novel poison para-aminopropiophenone (PAPP) which may present a hazard to some native animal populations. Here we used experimental and predictive approaches to evaluate feral cat bait take by a large native Australian predatory reptile the Lace monitor (Varanus varius). These lizards would be expected to readily detect, ingest and consume a lethal dose (depending on toxin) from surface-laid baits intended for feral cat control if a precautionary approach was not adopted when baiting. We modelled V. varius bait take using experimental and predictive biophysical modelling approaches to evaluate temporal effects of climate variables on V. varius activity and hence potential for bait removal. Finally we conducted a pre-PAPP baiting site occupancy assessment of V. varius within Wilson Promontory National Park (WPNP) to provide a basis for monitoring any longer term population effects of cat baiting. V. varius removed 7 % of deployed baits from 73 % of bait stations across another study area in Far Eastern Victoria. Daily bait removal was positively correlated with maximum temperature and solar radiation. Biophysical modelling for Far Eastern Victoria predicted that maximum temperatures <19.5 °C prevented V. varius activity and hence opportunity for bait removal. V. varius in WPNP was undetectable suggesting aerial baiting posed limited hazard to this species at this location. Depending how climate influences annual activity patterns and the specific poison, surface-laid baits could pose a significant mortality risk to V. varius. However, use of biophysical models to predict periods of V. varius inactivity may provide a novel means to reduce non-target bait take by this predator.     

Development of a bait and baiting system for delivery of oral rabies vaccine to free-ranging African wild dogs (Lycaon pictus)

The objective of the study was to develop a bait and baiting system capable of delivering one effective dose of oral rabies vaccine to each member of a free-ranging African wild dog (Lycaon pictus) pack. Trials were conducted between June and October 2000. The results of cafeteria-style bait preference trials testing seven candidate baits in captive wild dogs revealed a significant preference for chicken heads (June trials: P = 0.023, September trials: P = 0.021). Trials using a topical biomarker (rhodamine B) showed that chicken head baits were sufficiently chewed on most occasions to rupture the vaccine container. Free-ranging wild dogs and young pups ingested chicken head baits. Significant dominance of bait intake by a single individual was seen in four of six study packs and in the three packs in which an alpha pair could be distinguished, the dominant feeder was an alpha animal. Pattern of bait distribution and degree of satiation had no effect on pack coverage (proportion of pack ingesting at least one bait). Pack coverage was significantly related to trial number (r = 0.71, P < 0.001), with pack coverage increasing with increased exposure of the pack to the baits. During 46 hr of diurnal observations of free-ranging wild dogs only two baits were lost to non-target species. A baiting system for the oral vaccination of captive and free-ranging wild dogs is proposed.     

Development and efficacy of three poison baits against Blattella germanica under laboratory conditions

Bait formulations are widely used for controlling the German cockroach, but cockroaches increasingly come into aversion and resistance to bait formulations. To optimize baits and delay insecticides resistance to Blattella germanica, five insecticides with different mechanisms of action were selected to test the stomach toxicity of B. germanica, and twelve nutrient-rich foods were selected to test the palatability of B. germanica in the research. The results revealed that flufiprole and dinotefuran had a significant effect on the decrease of B. germanica population, whereas hydramethylnon, permethrin did not work well in 72 h stomach toxicity and acrinathrin had a short potency in pesticide formulation because of its semi-volatile physical properties. Furthermore, B. germanica prefers the poison bait to rat feed. This research represents the bait consisting of peanut powder and maltose (9:1) has the best palatability to B. germanica, and flufiprole and dinotefuran have a lot of potential in baits development and application.     

Factors Influencing Uptake of Sylvatic Plague Vaccine Baits by Prairie Dogs

Sylvatic plague vaccine (SPV) is a virally vectored bait-delivered vaccine expressing Yersinia pestis antigens that can protect prairie dogs (Cynomys spp.) from plague and has potential utility as a management tool. In a large-scale 3-year field trial, SPV-laden baits containing the biomarker rhodamine B (used to determine bait consumption) were distributed annually at a rate of approximately 100–125 baits/hectare along transects at 58 plots encompassing the geographic ranges of four species of prairie dogs. We assessed site- and individual-level factors related to bait uptake in prairie dogs to determine which were associated with bait uptake rates. Overall bait uptake for 7820 prairie dogs sampled was 70% (95% C.I. 69.9–72.0). Factors influencing bait uptake rates by prairie dogs varied by species, however, in general, heavier animals had greater bait uptake rates. Vegetation quality and day of baiting influenced this relationship for black-tailed, Gunnison’s, and Utah prairie dogs. For these species, baiting later in the season, when normalized difference vegetation indices (a measure of green vegetation density) are lower, improves bait uptake by smaller animals. Consideration of these factors can aid in the development of species-specific SPV baiting strategies that maximize bait uptake and subsequent immunization of prairie dogs against plague.     

Baiting studies on oral vaccination of the greater kudu (<Emphasis Type="Italic">Tragelaphus strepsiceros</Emphasis>) against rabies

Rabies in the greater kudu (Tragelaphus strepsiceros), one of the largest African antelopes, is a phenomenon unique to Namibia. Since the mid-1970s, the country has been plagued by two epizootics that claimed thousands of casualties among the indigenous kudu population. Reasons as to why kudus appear to be exceptionally susceptible to the disease still remain speculative at best. Because the Namibian economy relies heavily on trophy hunting, game meat hunting, and ecotourism, the current severe spread of rabies among kudus calls for effective measures to protect these antelopes. Oral vaccination of kudus may offer a possibility provided efficacious oral rabies vaccines are available. In this screening study, we explored options for attractive baits that facilitate optimal vaccine uptake by the target species and a bait distribution system that maximizes bait availability to the target species meanwhile minimizing bait depletion by non-target species. The results show that gelatin-based baits mixed with local (pods of the camel thorn tree) but also imported (apple-flavored corn meal) attractants are highly attractive for kudus providing a basis for future oral rabies vaccine baits.     

Acceptance of baits for delivery of oral rabies vaccine to raccoons.

During 1993-96 the Ontario Ministry of Natural Resources (Canada) implemented a research project to evaluate the efficacy of three candidate baits to deliver oral rabies vaccine to wild raccoons (Procyon lotor). Extensive field testing revealed that raccoon acceptance of Sugar-Vanilla baits (SV) at densities of 200/km2 and 400/km2, hand-placed in urban habitats of Scarborough (Ontario) during 1993, was 74% and 82%, respectively. Raccoon density in those areas averaged 11/km2. Aerial placement of SV baits in rural habitats in Barrie (Ontario) during 1993 and 1994, yielded raccoon acceptance levels of 58% with a density of 100 baits/km2, 59% at 75 baits/km2, and 47% at 50 baits/km2. Raccoon acceptance of SV baits was significantly lower in areas baited at the density of 50/km2. Acceptance of Cheese baits (CH) at a density of 75 baits/km2 was 52%. During 1996 trials in Barrie, modified SV baits with blister packs protruding through the matrix yielded raccoon acceptance values of 51% at a bait density of 54/km2, whereas acceptance of regular SV baits was 39% at a density of 51 baits/km2. Pooling of bait acceptance data for all years revealed that bait acceptance was highest for adult male raccoons. Raccoon density in rural habitats (Barrie, Ontario) where the studies took place, averaged 11-13/km2. Puncture and impact testing of blister packs in baits suggested that they would adequately serve as a vehicle to contain oral rabies vaccine for delivery to raccoons via baits.     

A New Recombinant BCG Vaccine Induces Specific Th17 and Th1 Effector Cells with Higher Protective Efficacy against Tuberculosis

Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (Mtb) that is a major public health problem. The vaccine used for TB prevention is Mycobacterium bovis bacillus Calmette-Guérin (BCG), which provides variable efficacy in protecting against pulmonary TB among adults. Consequently, several groups have pursued the development of a new vaccine with a superior protective capacity to that of BCG. Here we constructed a new recombinant BCG (rBCG) vaccine expressing a fusion protein (CMX) composed of immune dominant epitopes from Ag85C, MPT51, and HspX and evaluated its immunogenicity and protection in a murine model of infection. The stability of the vaccine in vivo was maintained for up to 20 days post-vaccination. rBCG-CMX was efficiently phagocytized by peritoneal macrophages and induced nitric oxide (NO) production. Following mouse immunization, this vaccine induced a specific immune response in cells from lungs and spleen to the fusion protein and to each of the component recombinant proteins by themselves. Vaccinated mice presented higher amounts of Th1, Th17, and polyfunctional specific T cells. rBCG-CMX vaccination reduced the extension of lung lesions caused by challenge with Mtb as well as the lung bacterial load. In addition, when this vaccine was used in a prime-boost strategy together with rCMX, the lung bacterial load was lower than the result observed by BCG vaccination. This study describes the creation of a new promising vaccine for TB that we hope will be used in further studies to address its safety before proceeding to clinical trials.     

Orchid (Orchidaceae) decline in the Catoctin Mountains,Frederick County,Maryland as documented by a long-term dataset

A 41-year study (1968–2008) of the orchids of the Catoctin Mountains, Frederick County, Maryland reveals that 19 of 21 species have experienced precipitous declines. Four of these species are currently considered Threatened or Endangered by the State of Maryland and another two are considered Rare. Annual census data at 167 sites from throughout the Catoctin Mountains on protected and unprotected lands (private and public) show a loss of three species from the study area, a decline of >90 % (ranging from 99 to 91 %) in seven species, and a decline of <90 % (ranging from 51 to 87 %) for nine species. Each species was analyzed using Ordinary Least Squares Analysis to show trends and document corresponding R² and p values. We tested the hypothesis that this decline is due to intensified herbivory by white-tailed deer. The overall orchid census data is significantly inversely-correlated (R = ?0.93) to the white-tailed deer harvest data of Frederick County (a surrogate for population size), which includes the entirety of the study area. Platanthera ciliaris showed a huge expansion at a single site explicitly managed for this species otherwise this orchid showed a decline similar to the other species. Proper management is critical for the continuation of the orchid species in this study, be it control of the white-tailed deer herd or combating woody plant succession in the case of P. ciliaris.