Seroprevalence, Blood Chemistry, and Patterns of Canine Parvovirus, Distemper Virus, Plague, and Tularemia in Free-Ranging Coyotes (Canis latrans) in Northern New Mexico, USA.

Wildlife diseases have implications for ecology, conservation, human health, and health of domestic animals. They may impact wildlife health and population dynamics. Exposure rates of coyotes (Canis latrans) to pathogens such as Yersinia pestis, the cause of plague, may reflect prevalence rates in both rodent prey and human populations. We captured coyotes in north-central New Mexico during 2005-2008 and collected blood samples for serologic surveys. We tested for antibodies against canine distemper virus (CDV, Canine morbillivirus), canine parvovirus (CPV, Carnivore protoparvovirus), plague, tularemia (Francisella tularensis), and for canine heartworm (Dirofilaria immitis) antigen. Serum biochemistry variables that fell outside reference ranges were probably related to capture stress. We detected antibodies to parvovirus in 32/32 samples (100%), and to Y. pestis in 26/31 (84%). More than half 19/32 (59%) had antibodies against CDV, and 5/31 (39%) had antibodies against F. tularensis. We did not detect any heartworm antigens (n = 9). Pathogen prevalence was similar between sexes and among the three coyote packs in the study area. Parvovirus exposure appeared to happen early in life, and prevalence of antibodies against CDV increased with increasing age class. Exposure to Y. pestis and F. tularensis occurred across all age classes. The high coyote seroprevalence rates observed for CPV, Y. pestis, and CDV may indicate high prevalence in sympatric vertebrate populations, with implications for regional wildlife conservation as well as risk to humans via zoonotic transmission.

Evaluation of a multi-species Protein A-ELISA assay for plague serologic diagnosis in humans and other mammal hosts.

BACKGROUND: The Hemagglutination assay (HA) is widely used in plague diagnosis, however, it has a subjective interpretation and demands high amounts of antigen and other immunobiological supplies. On the other hand, the conventional Anti-IgG ELISA is limited by the need of specific conjugates for multiple plague hosts, which leaves a gap for new diagnostic methods able to cover both the diagnosis of human cases and the epidemiological surveillance of multiple sentinel species. METHODS: We developed an ELISA Protein A-peroxidase method to detect anti-F1 antibodies across several species, including humans. To determine the cut-off and performance rates, HA results from 288 samples (81 rabbits, 64 humans, 66 rodents and 77 dogs) were used as reference. Next, we evaluated the agreement between Protein A-ELISA and Anti-IgG ELISA in an expanded sample set (n = 487). RESULTS: Optimal conditions were found with 250ng/well of F1 and 1:500 serum dilution. Protein A-ELISA showed high repeatability and reproducibility. We observed good correlation rates between the Protein A and IgG ELISAs optical densities and a higher positive/negative OD ratio for the Protein A-ELISA method. The overall sensitivity, specificity and area under the curve for Protein A-ELISA were 94%, 99% and 0.99, respectively. Similar results were observed for each species separately. In the analysis of the expanded sample set, there was a strong agreement between Protein A and IgG assays (kappa = 0.97). Furthermore, there was no cross-reaction with other common infectious diseases, such as dengue, Zika, Chagas disease, tuberculosis (humans) and ehrlichiosis, anaplasmosis and leishmaniasis (dogs). CONCLUSIONS: Altogether, the Protein A-ELISA showed high performance when compared both to HA and Anti-IgG ELISA, with a polyvalent single protocol that requires reduced amounts of antigen and can be employed to any plague hosts.

Performance assessment of a new indirect rapid diagnostic test for plague detection in humans and other mammalian hosts.

Plague is a flea-borne zoonosis that affects a wide range of mammals and still causes outbreaks in human populations yearly across several countries. While crucial for proper treatment, early diagnosis is still a major challenge in low- and middle-income countries due to poor access to laboratory infrastructure in rural areas. To tackle this issue, we developed and evaluated a new Fraction 1 capsular antigen (F1)-based rapid diagnostic test (RDT) as an alternative method for plague serological diagnosis and surveillance in humans and other mammals. In this study, 187 serum samples from humans, dogs, rodents and rabbits were retrospectively assessed using the plague RDT method. To calculate its performance, results were compared to those obtained by traditional hemagglutination (HA) and ELISA, which are well-established methods in the plague routine serodiagnosis. Remarkably, the results from RDT were in full agreement with those from the ELISA and HA assays, resulting in 100% (CI 95% = 95.5-100%) of sensitivity and 100% (CI 95% = 96.6-100%) of specificity. Accordingly, the Cohen's Kappa test coefficient was 1.0 (almost perfect agreement). Moreover, the RDT showed no cross-reaction when tested with sera from individuals positive to other pathogens, such as Y. pseudotuberculosis, Yersinia enterocolitica, Anaplasma platys, Ehrlichia canis and Leishmania infantum. Although preliminary, this study brings consistent proof-of-concept results with high performance of the Plague RDT when compared to HA and ELISA. Although further human and animal population-based studies will be necessary to validate these findings, the data presented here show that the plague RDT is highly sensitive and specific, polyvalent to several mammal species and simple to use in field surveillance or point-of-care situations with instant results.

An Evaluation of the Flea Index as a Predictor of Plague Epizootics in the West Nile Region of Uganda.

Plague is a low incidence flea-borne zoonosis that is often fatal if treatment is delayed or inadequate. Outbreaks occur sporadically and human cases are often preceded by epizootics among rodents. Early recognition of epizootics coupled with appropriate prevention measures should reduce plague morbidity and mortality. For nearly a century, the flea index (a measure of fleas per host) has been used as a measure of risk for epizootic spread and human plague case occurrence, yet the practicality and effectiveness of its use in surveillance programs has not been evaluated rigorously. We sought to determine whether long-term monitoring of the Xenopsylla flea index on hut-dwelling rats in sentinel villages in the plague-endemic West Nile region of Uganda accurately predicted plague occurrence in the surrounding parish. Based on observations spanning ~6 yr, we showed that on average, the Xenopsylla flea index increased prior to the start of the annual plague season and tended to be higher in years when plague activity was reported in humans or rodents compared with years when it was not. However, this labor-intensive effort had limited spatial coverage and was a poor predictor of plague activity within sentinel parishes.

Ongoing quiescence in the Borborema Plateau Plague focus (Paraiba, Brazil).

Plague is a zoonosis caused by Yersinia pestis, whose cycle is based on a reservoir system composed of mammals and their fleas. Its transmission cycle presents long enzootic periods with undetected cases, sometimes misleading that the cycle is extinct. While surveillance activities in Brazil are being carried out only in some focal areas, the serologic results confirm the persistence of Y. pestis in all monitored areas. We studied the small mammal assembly and Y. pestis presencein the Borborema Plateau Focus within the state of Paraíba, which staged the last Brazilian plague outbreak (1986-1987), through aninventory and Y. pestis detection survey of small mammals in peridomestic and sylvatic areas from two municipalities in the state of Paraíba.The field sampling captured 45 specimens (27 marsupials, 18 rodents), of 10 species. Only two species (one marsupial, one rodent) were captured in both peridomestic and sylvatic ecotopes. The sylvatic ecotope had higher richness and abundance. No evidence of circulation of the pathogen was detected, however, this result does not discard the necessity of continuous epidemiological surveillance due to the risk of rekindling the foci after long dormant periods, especially given the current epidemiological transition occurring on a Global scale.

Seroprevalence and spatial distribution dynamics of Yersinia pestis antibodies in dogs and cats from plague foci in the State of Ceará, Northeastern Brazil

Abstract INTRODUCTION: In Brazil, the plague is established in several foci located mainly in the northeastern part of the country, where it alternates between active and quiescent periods. These foci in the State of Ceará have high epidemiological importance. In addition to other plague detection activities, plague areas can be monitored through serological surveys of dogs and cats (domestic carnivores), which, following feeding on plague-infected rodents, can develop mild to severe forms of the disease and produce long-lasting antibodies. This study aimed to characterize the circulation dynamics and spatial distribution of Yersinia pestis antibodies in dogs and cats in plague foci areas of Ceará. METHODS: An ecological study was conducted to analyze the temporal series and spatial distribution of secondary data obtained from domestic carnivore serum surveillance in Ceará's plague areas from 1990 to 2014. RESULTS: Joinpoint analysis revealed that the overall trend was a reduction in antibody-positive animals. The mean proportion of antibody-positivity during the whole study period was 1.5% (3,023/203,311) for dogs, and 0.7% (426/61,135) for cats, with more than 4% antibody-positivity in dogs in 1997 and 2002. Antibody titers ranging from 1/16 to 1/64 were frequent. Despite fluctuations and a significant reduction, in recent years, there were antibody-positive animals annually throughout the study period, and the localities containing antibody-positive animals increased in number. CONCLUSION: Yersinia pestis is actively circulating in the study areas, posing a danger to the human population.

Seroprevalence and spatial distribution dynamics of Yersinia pestis antibodies in dogs and cats from plague foci in the State of Ceará, Northeastern Brazil.

INTRODUCTION: In Brazil, the plague is established in several foci located mainly in the northeastern part of the country, where it alternates between active and quiescent periods. These foci in the State of Ceará have high epidemiological importance. In addition to other plague detection activities, plague areas can be monitored through serological surveys of dogs and cats (domestic carnivores), which, following feeding on plague-infected rodents, can develop mild to severe forms of the disease and produce long-lasting antibodies. This study aimed to characterize the circulation dynamics and spatial distribution of Yersinia pestis antibodies in dogs and cats in plague foci areas of Ceará. METHODS: An ecological study was conducted to analyze the temporal series and spatial distribution of secondary data obtained from domestic carnivore serum surveillance in Ceará's plague areas from 1990 to 2014. RESULTS: Joinpoint analysis revealed that the overall trend was a reduction in antibody-positive animals. The mean proportion of antibody-positivity during the whole study period was 1.5% (3,023/203,311) for dogs, and 0.7% (426/61,135) for cats, with more than 4% antibody-positivity in dogs in 1997 and 2002. Antibody titers ranging from 1/16 to 1/64 were frequent. Despite fluctuations and a significant reduction, in recent years, there were antibody-positive animals annually throughout the study period, and the localities containing antibody-positive animals increased in number. CONCLUSION: Yersinia pestis is actively circulating in the study areas, posing a danger to the human population.

Plague bacterium as a transformer species in prairie dogs and the grasslands of western North America.

Invasive transformer species change the character, condition, form, or nature of ecosystems and deserve considerable attention from conservation scientists. We applied the transformer species concept to the plague bacterium Yersinia pestis in western North America, where the pathogen was introduced around 1900. Y. pestis transforms grassland ecosystems by severely depleting the abundance of prairie dogs (Cynomys spp.) and thereby causing declines in native species abundance and diversity, including threatened and endangered species; altering food web connections; altering the import and export of nutrients; causing a loss of ecosystem resilience to encroaching invasive plants; and modifying prairie dog burrows. Y. pestis poses an important challenge to conservation biologists because it causes trophic-level perturbations that affect the stability of ecosystems. Unfortunately, understanding of the effects of Y. pestis on ecosystems is rudimentary, highlighting an acute need for continued research.

La Bacteria de la Peste como una Especie Transformadora en los Perritos de las Praderas y los Pastizales del Oeste de Norteamérica Resumen Las especies invasoras transformadoras cambian el carácter, la condición, la forma o la naturaleza de los ecosistemas y merecen atención considerable por parte de los científicos de la conservación. Aplicamos el concepto de especie transformadora a la bacteria de la peste Yersinia pestis en el oeste de Norteamérica, en donde el patógeno fue introducido alrededor de 1900. Y. pestis transforma los ecosistemas de pastizal al disminuir severamente la abundancia de los perritos de las praderas (Cynomys spp.) y por lo tanto causa declinaciones en la abundancia y diversidad de las especies nativas, incluidas las especies amenazadas y en peligro; altera las conexiones de las redes alimenticias; altera la importación y exportación de nutrientes; causa la pérdida de resiliencia del ecosistema ante las plantas invasoras; y modifica las madrigueras de los perritos. Y. pestis es un reto importante para los biólogos de la conservación ya que causa perturbaciones de nivel trófico que afectan la estabilidad de los ecosistemas. Desafortunadamente, el entendimiento de los efectos de Y. pestis sobre los ecosistemas es rudimentario, lo que resalta una necesidad aguda de investigación continua.

Prevalence of the generalist flea Pulex simulans on black-tailed prairie dogs (Cynomys ludovicianus) in New Mexico, USA: the importance of considering imperfect detection.

If a parasite is not detected during a survey, one of two explanations is possible: the parasite was truly absent or it was present but not detected. We fit occupancy models to account for imperfect detection when combing fleas (Siphonaptera) from black-tailed prairie dogs (Cynomys ludovicianus) during June-August 2012 in the Vermejo Park Ranch, New Mexico, USA. With the use of detection histories from combing events during monthly trapping sessions, we fit occupancy models for two flea species: Oropsylla hirusta (a prairie dog specialist) and Pulex simulans (a generalist). Detection probability was <100% for both species and about 21% lower for P. simulans. Pulex simulans may be especially difficult to detect because it is about half the size of O. hirusta. Monthly occupancy (prevalence) for P. simulans was estimated at 24% (June, 95% confidence interval = 19-30), 39% (July, 32-47), and 56% (August, 49-64) in new prairie dog colonies, and 43% (32-54), 61% (49-71), and 79% (70-87) in old colonies. These results suggest P. simulans can attain high prevalence on prairie dogs, especially in old colonies. If P. simulans is highly prevalent on prairie dogs, it may serve as a "bridge vector" between Cynomys and other mammalian hosts of the plague bacterium Yersinia pestis, and even function as a reservoir of Y. pestis between outbreaks.

[Historical review of the plague in South America: a little-known disease in Colombia]. / Reseña histórica de la peste en Suramérica: una enfermedad poco conocida en Colombia.

The plague is an infectious disease that has transcended through history and has been responsible for three pandemics with high mortality rates. During the third pandemic that started in Hong Kong (1894), the disease spread through maritime routes to different regions in the world, including South America. In this region, approximately 16 million people are thought to be at risk in relation to this disease due to specific situations like human-rodent coexistence inside houses in rural areas, homes built with inadequate materials that are vulnerable to invasion by these animals, inappropriate storage of crops and an increase in rainfall and deforestation, which allows for the displacement of wild fauna and man invasion of the natural foci of the disease. Between 1994 and 1999, five countries: Bolivia, Brazil, Ecuador, Peru and the United States of America, reported approximately 1,700 cases with 79 related deaths. In Colombia we have historical data about an "infectious pneumonia" with high mortality rates that occurred during the same months, for three consecutive years (1913 to 1915) in the departments of Magdalena, Atlántico and Bolívar, located in the Colombian Atlantic coast, which suggested plague, but could not be confirmed.

Flea abundance, diversity, and plague in Gunnison's prairie dogs (Cynomys gunnisoni) and their burrows in montane grasslands in northern New Mexico.

Plague, a flea-transmitted infectious disease caused by the bacterium Yersinia pestis, is a primary threat to the persistence of prairie dog populations (Cynomys spp.). We conducted a 3-yr survey (2004-2006) of fleas from Gunnison's prairie dogs (Cynomys gunnisoni) and their burrows in montane grasslands in Valles Caldera National Preserve in New Mexico. Our objectives were to describe flea communities and identify flea and rodent species important to the maintenance of plague. We live-trapped prairie dogs and conducted burrow sweeps at three colonies in spring and summer of each year. One hundred thirty prairie dogs and 51 golden-mantled ground squirrels (Spermophilus lateralis) were captured over 3,640 trap nights and 320 burrows were swabbed for fleas. Five flea species were identified from prairie dogs and ground squirrels and four were identified from burrow samples. Oropsylla hirsuta was the most abundant species found on prairie dogs and in burrows. Oropsylla idahoensis was most common on ground squirrels. Two colonies experienced plague epizootics in fall 2004. Plague-positive fleas were recovered from burrows (O. hirsuta and Oropsylla tuberculata tuberculata) and a prairie dog (O. hirsuta) in spring 2005 and summer 2006. Three prairie dogs collected in summer 2005 and 2006 had plague antibody. We found a significant surge in flea abundance and prevalence, particularly within burrows, following plague exposure. We noted an increased tendency for flea exchange opportunities in the spring before O. hirsuta reached its peak population. We hypothesize that the role of burrows as a site of flea exchange, particularly between prairie dogs and ground squirrels, may be as important as summer conditions that lead to buildup in O. hirsuta populations for determining plague outbreaks.

Prevalência de anticorpos contra Yersinia pestis em carnívoros domésticos nos focos pestosos, do Estado do Ceará / Prevalence of antibodies against Yersinia pestis in domestic carnivores, in plague foci in the State of Ceará

Analisou-se a prevalência de anticorpos contra Yersinia pestis em carnívoros domésticos (cães e gatos) nas áreas pestígenas do Estado do Ceará, visando estabelecer a importância do monitoramento desses animais na rotina do Programa de Controle da Peste. No decênio 1997-2006, analisaram-se 146.732 amostras de soros (95.883 cães e 50.849 gatos), das quais 2.629 (2.234 cães e 395 gatos) revelaram-se positivas. A prevalência entre os cães (85 por cento) foi superior a dos gatos (15 por cento) em todo o decênio e locais, exceto em Ibiapina, em 1998. O significado desses achados ainda não foi determinado. Os estudos sobre a zoonose no Brasil pautaram-se por paradigmas que não contemplavam todos os elementos envolvidos na zoonose, impossibilitando a devida elucidação do papel desses carnívoros. O monitoramento da atividade pestosa, realizado exclusivamente por inquéritos caninos, pode redundar no desconhecimento progressivo da situação epidemiológica da peste, caso não sejam desenvolvidas pesquisas interinstitucionais suplementares.

The prevalence of antibodies against Yersinia pestis in domestic carnivores (dogs and cats), in plague areas in the State of Ceará, was analyzed to establish the importance of monitoring these animals within the routine practice of the plague control program. Over the decade 1997-2006, 146,732 serum samples were examined (95,883 from dogs and 50,849 from cats), of which 2,629 (2,234 from dogs and 395 from cats) proved to be positive. The prevalence among dogs (85 percent) was higher than among cats (15 percent) throughout the decade and in all places, except in Ibiapina in 1998. The significance of these findings has not yet been determined. Studies on this zoonosis in Brazil have been based on paradigms that did not cover all the elements involved in the zoonosis, thus making it impossible to properly understand the role of these carnivores. Monitoring of plague foci conducted exclusively by means of dog surveys may result in progressive lack of knowledge of the epidemiological situation of plague, if supplementary inter-institutional research is not developed.

[Prevalence of antibodies against Yersinia pestis in domestic carnivores, in plague foci in the State of Ceará]. / Prevalência de anticorpos contra Yersinia pestis em carnívoros domésticos nos focos pestosos, do Estado do Ceará

The prevalence of antibodies against Yersinia pestis in domestic carnivores (dogs and cats), in plague areas in the State of Ceará, was analyzed to establish the importance of monitoring these animals within the routine practice of the plague control program. Over the decade 1997-2006, 146,732 serum samples were examined (95,883 from dogs and 50,849 from cats), of which 2,629 (2,234 from dogs and 395 from cats) proved to be positive. The prevalence among dogs (85%) was higher than among cats (15%) throughout the decade and in all places, except in Ibiapina in 1998. The significance of these findings has not yet been determined. Studies on this zoonosis in Brazil have been based on paradigms that did not cover all the elements involved in the zoonosis, thus making it impossible to properly understand the role of these carnivores. Monitoring of plague foci conducted exclusively by means of dog surveys may result in progressive lack of knowledge of the epidemiological situation of plague, if supplementary inter-institutional research is not developed.

[Surveillance of plague in the State of Ceará: 1990-1999]. / Vigilância da peste no Estado do Ceará: 1990-1999.

Serological surveillance activities regarding the foci of plague in Ceará State have detected a rising number of sentinel animals with antiplague antibodies in 1995, with a peak in 1997 demonstrating an increase in the plague bacteria activities throughout all the foci investigated. From a total of 110,725 serum samples collected from rodents (7,873) and domestic carnivores (102,852) analyzed by the Hemaglutination technique (HA) for antibodies against F1 antigen of Yersinia pestis 905 samples tested positive. In these samples there were 15 rodents (4 Rattus rattus and 11 Galea spp), 720 dogs and 170 cats. Of the 652 human suspected and contact cases investigated by HA, only two were positive. A third case had a positive hemoculture for Y. pestis. The isolate is highly pathogenic for laboratory animals and showed sensitivity to the antimicrobial drugs used for plague treatment.

Vigilância da peste no Estado do Ceará: 1990-1999 / Surveillance of plague in the State of Ceará: 1990-1999

As atividades de vigilância sorológica da peste nos focos do Estado do Ceará detectaram elevaçäo do número de animais indicadores/sentinela com anticorpos antipestosos a partir de 1995, com pico em 1997 evidenciando aumento da circulaçäo do bacilo pestoso em todos os focos investigados. De um total de 110.725 amostras de soro obtidas de roedores (7.873) e carnívoros domésticos (102.852) analisadas pela técnica de hemaglutinaçäo (HA) para detecçäo de anticorpos contra o antígeno F1 da Yersinia pestis, 905 revelaram-se positivas, sendo 15 de roedores (4 Rattus rattus e 11 Galea spp), 720 de cäes e 170 de gatos. Dos 652 casos humanos suspeitos e contatos investigados apenas dois foram positivos pela HA e um terceiro paciente foi positivo por hemocultura. A cepa isolada revelou-se altamente patogênica para animais de laboratório e mostrou sensibilidade aos antimicrobianos usados no tratamento dos doentes

Dynamics of plague in a Gunnison's prairie dog colony complex from New Mexico.

A plague (Yersinia pestis) epizootic spread through Gunnison's prairie dogs (Cynomys gunnisoni), and possibly other rodent species, in the Moreno Valley in north-central New Mexico between winter 1984-1985 and autumn 1987. We observed the progress of the epizootic and subsequent population recovery at four prairie dog towns within the valley during this period. At two towns (Midlake and Val Verde) the prairie dogs were marked prior to the epizootic. At two additional towns (Vega and South Entrance) prairie dogs were marked following the epizootic. In 1988, a second epizootic occurred at Vega. One hundred thirty-nine serum samples were collected from prairie dogs and other rodents and 1,750 fleas were collected from animals and burrows. Fleas infected with Y. pestis were collected from prairie dogs, deer mice (Peromyscus maniculatus), and thirteen-lined ground squirrels (Spermophilus tridecemlineatus). Prairie dog fleas included Oropsylla hirsuta, O. labis and O. tuberculata, deermouse associated fleas were Aetheca wagneri and Rhadinopsylla sectilis, and Oropsylla bacchi was associated with thirteen-lined ground squirrels. All of the above flea species were collected from prairie dog burrows. All rodent species shared some flea species. Thirteen-lined ground squirrels disappeared shortly before plague was identified in prairie dogs at Midlake. Meadow voles were rare following the epizootic at Vega in 1986, became abundant in 1987, and disappeared at the time of the second prairie dog epizootic in summer 1988. Although we collected serum from Gunnison's prairie dogs, thirteen-lined ground squirrels, deer mice, and meadow voles (Microtus pennsylvanicus), we identified elevated serum titers against Y. pestis only in Gunnison's prairie dogs. Prairie dog mortality at all towns affected by plague was in excess of 99%. Serum antibody titers indicate that more than 40% of the few prairie dogs left to establish colonies following epizootics survived plague infection.