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Effects of experimental flea removal and plague vaccine treatments on survival of northern Idaho ground squirrels and two coexisting sciurids

Plague is a non-native disease in North America that reduces survival of many mammals. Previous studies have focused on epizootic plague which causes acute mortality events and dramatic declines in local abundance. We know much less about enzootic plague which causes less punctuated reductions in survival and abundance of infected populations. As a result, enzootic plague is much more difficult to detect because changes in population attributes are more subtle and Yersinia pestis prevalence is likely lower relative to epizootic plague outbreaks. The northern Idaho ground squirrel ( Urocitellus brunneus ) is a threatened species which coexists with Columbian ground squirrels ( Urocitellus columbianus ) and yellow-pine chipmunks ( Neotamias amoenus ) throughout their restricted distribution in central Idaho. Columbian ground squirrels and yellow-pine chipmunks are more abundant and widespread than northern Idaho ground squirrels and both are known hosts for plague. Hence, enzootic plague may be one cause of rarity for northern Idaho ground squirrels but its effect on this threatened species has not been evaluated. We conducted three controlled and randomized field experiments to examine the effects of plague in northern Idaho ground squirrels and the two coexisting species: 1) a plague vaccine experiment, 2) a paired flea-reduction experiment, and 3) a non-paired flea-reduction experiment. For Experiment 1, we hypothesized that if enzootic plague is present, vaccinated animals would have higher survival. Furthermore, Experiments 2 and 3 tested the prediction that untreated, control animals should have lower survival than those in areas where fleas are experimentally removed or reduced because fleas are the main vector for plague. In the plague vaccine experiment, vaccinated chipmunks had 4.65% higher apparent survival compared to chipmunks that received a placebo for intervals when the vaccine is believed to be effective. Apparent annual survival increased for all three species on experimental flea-reduction plots compared to non-treated plots for the paired experiment but results were mixed for the non-paired experiment. Taken together, our results suggest that enzootic plague is present and negatively impacting survival of northern Idaho ground squirrels and two coexisting species.

Idaho

Virally-vectored vaccine candidates against white-nose syndrome induce anti-fungal immune response in little brown bats (Myotis lucifugus)

White-nose syndrome (WNS) caused by the fungus, Pseudogymnoascus destructans ( Pd ) has killed millions of North American hibernating bats. Currently, methods to prevent the disease are limited. We conducted two trials to assess potential WNS vaccine candidates in wild-caught Myotis lucifugus . In a pilot study, we immunized bats with one of four vaccine treatments or phosphate-buffered saline (PBS) as a control and challenged them with Pd upon transfer into hibernation chambers. Bats in one vaccine-treated group, that received raccoon poxviruses (RCN) expressing Pd calnexin (CAL) and serine protease (SP), developed WNS at a lower rate (1/10) than other treatments combined (14/23), although samples sizes were small. The results of a second similar trial provided additional support for this observation. Bats vaccinated orally or by injection with RCN-CAL and RCN-SP survived Pd challenge at a significantly higher rate (P = 0.01) than controls. Using RT-PCR and flow cytometry, combined with fluorescent in situ hybridization, we determined that expression of IFN-γ transcripts and the number of CD4 + T-helper cells transcribing this gene were elevated (P < 0.10) in stimulated lymphocytes from surviving vaccinees (n = 15) compared to controls (n = 3). We conclude that vaccination with virally-vectored Pd antigens induced antifungal immunity that could potentially protect bats against WNS.

Scientific Reports

Vaccination of rainbow trout against infectious hematopoietic necrosis (IHN) by using attenuated mutants selected by neutralizing monoclonal antibodies

A neutralizing monoclonal antibody against infectious hematopoietic necrosis virus (IHNV) was used to select neutralization-resistant mutants from isolates of virus obtained from adult steelhead Oncorhynchus mykiss returning to the Round Butte Hatchery (RB mutants) on the Deschutes River in Oregon, USA, and from rainbow trout (nonanadromous O. mykiss) at a commercial hatchery in the Hagerman Valley of Idaho, USA (193-110 mutants). Two of the mutants, RB-1 and 193-110-4, were significantly (P < 0.001) attenuated compared with parental strains. Vaccination of rainbow trout by waterborne exposure to the mutants conferred solid protection against challenge with wild-type virus. In some trials, fish vaccinated with the RB-1 mutant at 50% tissue culture infectious doses (TCID50) of 1 X 104-1 x 105 TCID50/mL or with the 193-110-4 mutant at 1 x 102-1 x 103 TCID50/mL, held for 14 d, then challenged with the homologous wild-type strain at 1 x 105 TCID50/mL showed relative percent survival of 95-100% (P < 0.005). There was no significant difference (P > 0.05) in protection among fish exposed to the RB-1 vaccine strain at a dose of 1 x 105 TCID50/mL for periods of either 1, 12, or 24 h, held for 14 d, and then challenged with the wild-type RB isolate, although the 1-h exposure seemed to be somewhat less effective. Fish were vaccinated with the RB-1 strain at 1 x 103-1 x 105 TCID50/mL for 24 h then challenged after 1, 7, 14, or 21 d with the wild-type RB isolate. No significant (P > 0.1) protection was observed at 1 d postvaccination, but the relative percent survival increased progressively at each subsequent challenge period, becoming statistically significant by day 7 (P < 0.001) and beyond. These results suggested that resistance to challenge with wild-type virus resulted from development of IHNV-specific immunity and not from viral interference or interferon induction, and they reinforce the potential of an attenuated vaccine to control this important disease.

Journal of Aquatic Animal Health

Antibody response to rabies vaccination in captive and freeranging wolves (Canis lupus)

Fourteen captive and five free-ranging Minnesota gray wolves (Canis lupus) were tested for the presence of rabies virus neutralizing antibodies (RVNA) after vaccination with an inactivated canine rabies vaccine. Blood was collected from all wolves prior to vaccination and at 1 mo postvaccination (PV) and from all captive and three wild wolves at 3 mo PV. In addition, one free-ranging wolf was sampled at 4 mo PV, and two free-ranging wolves were sampled at 6 mo PV. All wolves were seronegative prior to vaccination. RVNA were detected in 14 (100%) captive wolves and in four of five (80%) free-ranging wolves. The geometric mean titer of the captive wolves at 1 mo PV was significantly higher (P = 0.023) than in the free-ranging wolves. Five of 13 (38.5%) captive wolves and none of the three (0%) free-ranging wolves had measurable RVNA at 3 mo PV. No measurable RVNA were detected in the serum samples collected from the free-ranging wolves at 4 and 6 mo PV. These results should be interpreted with caution because of the small number of free-ranging wolves tested. Further research is needed to properly assess immune function and antibody response to vaccination in captive wolves in comparison with their free-ranging counterparts.

Journal of Zoo and Wildlife Medicine

Vaccination as a potential means to prevent plague in black-footed ferrets: Progress and continuing challenges

This study was conducted to further assess the feasibility of vaccinating black-footed ferrets ( Mustela nigripes ) against plague (caused by the bacterium Yersinia pestis ). On days 0 and 28, 17 postreproductive ferrets were immunized by subcutaneous injection with a recombinant fusion protein containing F1 and V antigens from Y. pestis. Another 17 animals received a placebo by the same route. Two weeks after the second immunization, mean antibody titers to Y. pestis F1 and V antigens were measured and found to be significantly higher in vaccinates than their preimmunization values (P < 0.0001) and significantly higher than the control values (P < 0.0001). Six months postimmunization, 16 vaccinates and eight controls were challenged with approximately 8,000 colony forming units of virulent plague by subcutaneous inoculation. Eleven of 16 vaccinates (69 percent) survived with no ill effects whereas all eight control animals died within 3a??6 days. Two months later, the 11 surviving vaccinates were challenged again by ingestion of a plague-infected mouse. None of the animals showed any ill effects and all survived. In contrast, seven control ferrets fed infected mice died within 2a??4 days, including one animal that did not actually ingest the mouse but was likely exposed to it. This study demonstrates that immunization of ferrets with the recombinant F1-V fusion protein can induce significant antibody responses and reduce their susceptibility to plague infection.

Scientific Investigations Report

A cohabitation challenge to compare the efficacies of vaccines for bacterial kidney disease (BKD) in chinook salmon Oncorhynchus tshawytscha

The relative efficacies of 1 commercial and 5 experimental vaccines for bacterial kidney disease (BKD) were compared through a cohabitation waterborne challenge. Groups of juvenile chinook salmon Oncorhynchus tshawytscha were vaccinated with one of the following: (1) killed Renibacterium salmoninarum ATCC 33209 (Rs 33209) cells; (2) killed Rs 33209 cells which had been heated to 37??C for 48 h, a process that destroys the p57 protein; (3) killed R. salmoninarum MT239 (Rs MT239) cells; (4) heated Rs MT239 cells; (5) a recombinant version of the p57 protein (r-p57) emulsified in Freund's incomplete adjuvant (FIA); (6) the commercial BKD vaccine Renogen; (7) phosphate-buffered saline (PBS) emulsified with an equal volume of FIA; or (8) PBS alone. Following injection, each fish was marked with a subcutaneous fluorescent latex tag denoting its treatment group and the vaccinated fish were combined into sham and disease challenge tanks. Two weeks after these fish were vaccinated, separate groups of fish were injected with either PBS or live R. salmoninarum GL64 and were placed inside coated-wire mesh cylinders (liveboxes) in the sham and disease challenge tanks, respectively. Mortalities in both tanks were recorded for 285 d. Any mortalities among the livebox fish were replaced with an appropriate cohort (infected with R. salmoninarum or healthy) fish. None of the bacterins evaluated in this study induced protective immunity against the R. salmoninarum shed from the infected livebox fish. The percentage survival within the test groups in the R. salmoninarum challenge tank ranged from 59% (heated Rs MT239 bacterin) to 81 % (PBS emulsified with FIA). There were no differences in the percentage survival among the PBS-, PBS/FIA-, r-p57-and Renogen-injected groups. There also were no differences in survival among the bacterin groups, regardless of whether the bacterial cells had been heated or left untreated prior to injection. ?? Inter-Research 2005.

Diseases of Aquatic Organisms

Immunogenicity, safety, and anti-viral efficacy of a subunit SARS-CoV-2 vaccine candidate in captive black-footed ferrets (Mustela nigripes) and their susceptibility to viral challenge

A preliminary vaccination trial against the emergent pathogen, SARS-CoV-2, was completed in captive black-footed ferrets ( Mustela nigripes; BFF) to assess safety, immunogenicity, and anti-viral efficacy. Vaccination and boosting of 15 BFF with purified SARS-CoV-2 S1 subunit protein produced a nearly 150-fold increase in mean antibody titers compared to pre-vaccination titers. Serum antibody responses were highest in young animals, but in all vaccinees, antibody response declined rapidly. Anti-viral activity from vaccinated and unvaccinated BFF was determined in vitro, as well as in vivo with a passive serum transfer study in mice. Transgenic mice that received BFF serum transfers and were subsequently challenged with SARS-CoV-2 had lung viral loads that negatively correlated ( p < 0.05) with the BFF serum titer received. Lastly, an experimental challenge study in a small group of BFF was completed to test susceptibility to SARS-CoV-2. Despite viral replication and shedding in the upper respiratory tract for up to 7 days post-challenge, no clinical disease was observed in either vaccinated or naive animals. The lack of morbidity or mortality observed indicates SARS-CoV-2 is unlikely to affect wild BFF populations, but infected captive animals pose a potential risk, albeit low, for humans and other animals.

Viruses

Season and application rates affect vaccine bait consumption by prairie dogs in Colorado and Utah, USA

Plague, a zoonotic disease caused by the bacterium Yersinia pestis , causes high rates of mortality in prairie dogs ( Cynomys spp.). An oral vaccine against plague has been developed for prairie dogs along with a palatable bait to deliver vaccine and a biomarker to track bait consumption. We conducted field trials between September 2009 and September 2012 to develop recommendations for bait distribution to deliver plague vaccine to prairie dogs. The objectives were to evaluate the use of the biomarker, rhodamine B, in field settings to compare bait distribution strategies, to compare uptake of baits distributed at different densities, to assess seasonal effects on bait uptake, and to measure bait uptake by nontarget small mammal species. Rhodamine B effectively marked prairie dogs' whiskers during these field trials. To compare bait distribution strategies, we applied baits around active burrows or along transects at densities of 32, 65, and 130 baits/ha. Distributing baits at active burrows or by transect did not affect uptake by prairie dogs. Distributing baits at rates of &ge;65/ha (or &ge;1 bait/active burrow) produced optimal uptake, and bait uptake by prairie dogs in the autumn was superior to uptake in the spring. Six other species of small mammals consumed baits during these trials. All four species of tested prairie dogs readily consumed the baits, demonstrating that vaccine uptake will not be an obstacle to plague control via oral vaccination.

Colorado, Utah

Managing prairie dogs by managing plague: a vaccine for the future?

The Black-footed Ferret Recovery Implementation Team Executive Committee is conducting a project to develop, and (hopefully) eventually implement, a plague vaccination program for prairie dogs. The project is a component of the Western Association of Fish and Wildlife Agencies Grasslands Conservation Initiative. An effective, field-worthy vaccine against plague could be the biggest breakthrough in recovery efforts for the black-footed ferret since the 1981 rediscovery of wild ferrets near Meeteetse, Wyoming. If proven efficacious, the vaccine could help agencies and stakeholder cooperators maintain specific populations of prairie dogs at robust levels, thus enhancing range-wide conservation of those species, as well recovery of the ferret, while enabling control of other prairie dog populations to resolve site-specific agricultural and human health concerns. The results of laboratory and field-testing in the early stages of developing this vaccine are preliminary but mostly encouraging. A plan for broad-scale application is being developed for possible use when testing has been completed and (if warranted) the vaccine is registered for governmental use. An overview of all aspects of the project is discussed.

Conference Paper

A recombinant raccoon poxvirus vaccine expressing both Yersinia pestis F1 and truncated V antigens protects animals against lethal plague.

In previous studies, we demonstrated in mice and prairie dogs that simultaneous administration of two recombinant raccoon poxviruses ( r RCN) expressing Yersinia pestis antigens (F1 and V307-a truncated version of the V protein) provided superior protection against plague challenge compared to individual single antigen constructs. To reduce costs of vaccine production and facilitate implementation of a sylvatic plague vaccine (SPV) control program for prairie dogs, a dual antigen construct is more desirable. Here we report the construction and characterization of a novel RCN-vectored vaccine that simultaneously expresses both F1 and V307 antigens. This dual antigen vaccine provided similar levels of protection against plague in both mice and prairie dogs as compared to simultaneous administration of the two single antigen constructs and was also shown to protect mice against an F1 negative strain of Y. pestis. . The equivalent safety, immunogenicity and efficacy profile of the dual RCN-F1/V307 construct warrants further evaluation in field efficacy studies in sylvatic plague endemic areas.

Vaccines

Evaluation of Francisella orientalis ΔpdpA as a live attenuated vaccine against piscine Francisellosis in Nile tilapia

Francisella orientalis is an important bacterial pathogen of marine and freshwater fish with worldwide distribution. Fish francisellosis is a severe subacute to chronic granulomatous disease, with high mortalities and high infectivity rates in cultured and wild fish. To date, there is no approved vaccine for this disease. In this study, we evaluated the efficacy of a defined F. orientalis pathogenicity determinant protein A ( pdpA ) mutant (Δ pdpA ) as a live attenuated immersion vaccine against subsequent immersion challenge with the wild-type organism. Immunized Nile tilapia Oreochromis niloticus were protected (45% relative percent survival) from the lethal challenges and presented significantly lower mortality than nonvaccinated and challenged treatments. Although serum IgM was significantly higher in immunized fish, similar bacterial loads were detected in vaccinated and nonvaccinated survivors. In conclusion, although the F. orientalis Δ pdpA is attenuated and effectively stimulated an adaptive immune response, the low relative percent survival and high bacterial persistence in survivors of immunized and challenged treatments indicates low suitability of Δ pdpA as a mucosal vaccine for tilapia under conditions used in this study.

Journal of Aquatic Animal Health

Vaccine effects on heterogeneity in susceptibility and implications for population health management

Heterogeneity in host susceptibility is a key determinant of infectious disease dynamics but is rarely accounted for in assessment of disease control measures. Understanding how susceptibility is distributed in populations, and how control measures change this distribution, is integral to predicting the course of epidemics with and without interventions. Using multiple experimental and modeling approaches, we show that rainbow trout have relatively homogeneous susceptibility to infection with infectious hematopoietic necrosis virus and that vaccination increases heterogeneity in susceptibility in a nearly all-or-nothing fashion. In a simple transmission model with an R 0 of 2, the highly heterogeneous vaccine protection would cause a 35 percentage-point reduction in outbreak size over an intervention inducing homogenous protection at the same mean level. More broadly, these findings provide validation of methodology that can help to reduce biases in predictions of vaccine impact in natural settings and provide insight into how vaccination shapes population susceptibility.

mBio

Immersion vaccination of sockeye salmon (Oncorhynchus nerka) with two pathogenic strains of Vibrio anguillarum

Sockeye salmon ( Oncorhynchus nerka ) were immersion-vaccinated in suspensions containing 5&ensp;&times;&ensp;10 7 , 5&ensp;&times;&ensp;10 6 , 5&ensp;&times;&ensp;10 5 , or 5&ensp;&times;&ensp;10 4 bacteria/mL of bivalent or monovalent, formalin-killed Vibrio anguillarum , Types I and II. The fish were split into two lots and held for 54&ensp;d. At that time one lot was challenged with living, virulent V . anguillarum , Type I, and one with living, virulent V . anguillarum , Type II. Immunization with bivalent bacterin effectively protected the fish from vibriosis, but monovalent vaccine was effective only against the homologous challenge. Immunization with the highest concentration of Type I monovalent bacterin resulted in 0% Type I and 58% Type II challenge mortality. Immunization with the highest concentration of Type II monovalent bacterin resulted in 41% Type I and 0% Type II challenge mortality. Immunization with the highest concentration of bivalent Type I/Type II bacterin resulted in 2% mortality in both challenges. Protective bacterins were effective at concentrations down to 5&ensp;&times;&ensp;10 5 bacteria/mL. Key words : immersion vaccination, bivalent vaccines, Vibrio anguillarum , vibriosis.

Journal of the Fisheries Research Board of Canada

A missing dimension in measures of vaccination impacts

Immunological protection, acquired from either natural infection or vaccination, varies among hosts, reflecting underlying biological variation and affecting population-level protection. Owing to the nature of resistance mechanisms, distributions of susceptibility and protection entangle with pathogen dose in a way that can be decoupled by adequately representing the dose dimension. Any infectious processes must depend in some fashion on dose, and empirical evidence exists for an effect of exposure dose on the probability of transmission to mumps-vaccinated hosts [1] , the case-fatality ratio of measles [2] , and the probability of infection and, given infection, of symptoms in cholera [3] . Extreme distributions of vaccine protection have been termed leaky (partially protects all hosts) and all-or-nothing (totally protects a proportion of hosts) [4] . These distributions can be distinguished in vaccine field trials from the time dependence of infections [5] . Frailty mixing models have also been proposed to estimate the distribution of protection from time to event data [6] , [7] , although the results are not comparable across regions unless there is explicit control for baseline transmission [8] . Distributions of host susceptibility and acquired protection can be estimated from dose-response data generated under controlled experimental conditions [9] – [11] and natural settings [12] , [13] . These distributions can guide research on mechanisms of protection, as well as enable model validity across the entire range of transmission intensities. We argue for a shift to a dose-dimension paradigm in infectious disease science and community health.

PLoS Pathogens

Modeling of future COVID-19 cases, hospitalizations, and deaths, by vaccination rates and nonpharmaceutical intervention scenarios — United States, April–September 2021

What is already known about this topic? Increases in COVID-19 cases in March and early April occurred despite a large-scale vaccination program. Increases coincided with the spread of SARS-CoV-2 variants and relaxation of nonpharmaceutical interventions (NPIs). What is added by this report? Data from six models indicate that with high vaccination coverage and moderate NPI adherence, hospitalizations and deaths will likely remain low nationally, with a sharp decline in cases projected by July 2021. Lower NPI adherence could lead to substantial increases in severe COVID-19 outcomes, even with improved vaccination coverage. What are the implications for public health practice? High vaccination coverage and compliance with NPIs are essential to control COVID-19 and prevent surges in hospitalizations and deaths in the coming months.

Morbidity and Mortality Weekly Report

Sylvatic plague vaccine and management of prairie dogs

Scientists at the USGS National Wildlife Health Center (NWHC), in collaboration with colleagues at the University of Wisconsin (UW), have developed a sylvatic plague vaccine that shows great promise in protecting prairie dogs against plague (Mencher and others, 2004; Rocke and others, 2010). Four species of prairie dogs reside in the United States and Canada, and all are highly susceptible to plague and regularly experience outbreaks with devastating losses. Along with habitat loss and poisoning, plague has contributed to a significant historical decline in prairie dog populations. By some estimates, prairie dogs now occupy only 1 to 2 percent of their former range (Proctor and others, 2006), with prairie dog colonies being now much smaller and fragmented than they were historically, making individual colonies more vulnerable to elimination by plague (Antolin and others, 2002). At least one species, the Utah prairie dog ( Cynomys parvidens ) is listed by the U.S. Fish and Wildlife Service (FWS) as "threatened." Controlling plague is a vital concern for ongoing management and conservation efforts for prairie dogs. Current efforts to halt the spread of plague in prairie dog colonies typically rely on dusting individual prairie dog burrows with pesticides to kill plague-infected fleas. Although flea-control insecticides, such as deltamethrin, are useful in stopping plague outbreaks in these prairie dog colonies, dusting of burrows is labor intensive and time consuming and may affect other insects and arthropods. As an alternative approach, NWHC and UW scientists developed a sylvatic plague vaccine (SPV) for prairie dogs that can be delivered via oral bait. Laboratory studies have shown that consumption of this vaccine-laden bait by different prairie dog species results in significant protection against plague infection that can last for at least 9 months (Rocke and others, 2010; Rocke, unpublished). Work has now shifted to optimizing baits and distribution methods for field delivery of the vaccine. Ultimately, the bait will be formulated in a size and shape that facilitates distribution by plane or overland vehicle. Field studies to assess the safety and efficacy of SPV are being planned. These studies will require involvement from numerous partners, including state and federal land management agencies, tribal organizations, private landowners and non-government agencies.

Fact Sheet

Use of Rhodamine B as a biomarker for oral plague vaccination of prairie dogs

Oral vaccination against Yersinia pestis could provide a feasible approach for controlling plague in prairie dogs ( Cynomys spp.) for conservation and public health purposes. Biomarkers are useful in wildlife vaccination programs to demonstrate exposure to vaccine baits. Rhodamine B (RB) was tested as a potential biomarker for oral plague vaccination because it allows nonlethal sampling of animals through hair, blood, and feces. We found that RB is an appropriate marker for bait uptake studies of C. ludovicianus) when used at concentrations <0.5% of bait mass dosed to deliver >10 mg RB per kg target animal mass. Whiskers with follicles provided the best sample for RB detection.

Journal of Wildlife Diseases

Development of a subunit vaccine for infectious pancreatic necrosis virus using a baculovirus insect/larvae system

Various attempts to develop a vaccine against infectious pancreatic necrosis virus (IPNV) have not yielded consistent results. Thus, at present, no commercial vaccine is available that can be used with confidence to immunize fry of salmon and trout. We generated a cDNA clone of the large genome segment A of an IPNV Sp strain and expressed all structural protein genes in insect cells and larvae using a baculovirus expression system. Green fluorescent protein was also co-expressed as a reporter molecule. High yields of IPNV proteins were obtained and the structural proteins self assembled to form virus-like particles (VLPs). We tested the immunogenicity of the putative VLP antigen in immersion vaccine experiments (two concentrations) in rainbow trout (Oncorhynchus mykiss) fry, and by intraperitoneal immunisation of Atlantic salmon (Salmo salar) pre-smolts using an oil adjuvant formulation. Rainbow trout were challenged by immersion using either the Sp or the VR-299 strain of IPNV two or three weeks post-vaccination, while Atlantic salmon were bath challenged with Sp strain after two months, after parr-smolt transformation. In the rainbow trout fry challenged two weeks post-immunization, cumulative mortality rates three weeks post challenge were 14 % in the fry that had received the highest dose versus 8 % in the control groups. No indication of protection was seen in repeated trials using a lower dose of antigen and challenge three weeks post-immunisation. The cumulative mortality rate of intraperitoneally immunised Atlantic salmon post-smolts four weeks post challenge was lower (56 %) than in the control fish (77 %), showing a dose-response pattern.

Conference Paper