Actinomycosis in a gray four-eyed opossum (Philander opossum) caused by a novel species of Schaalia.

BACKGROUND: Infective lesions of the jaws and adjacent tissues (lumpy jaw disease, LJD) have been recognized as one major cause of death of captive macropods. Fusobacterium necrophorum and Actinomyces species serve as the main source of LJD in kangaroos and wallabies. Currently, little is reported about LJD or similar diseases in opossums. CASE PRESENTATION: Here we report a case of actinomycosis resembling the entity lumpy jaw disease in a gray four-eyed opossum, caused by a novel species of Schaalia. A 2.8 year old male Philander opossum was presented with unilateral swelling of the right mandible. After an initial treatment with marbofloxacin, the opossum was found dead the following day and the carcass was submitted for necropsy. Postmortem examination revealed severe mandibular skin and underlying soft tissue infection with subsequent septicemia as the cause of death. Histological examination demonstrated Splendore-Hoeppli phenomenon, typically seen in classical cases of actinomycosis. Bacteriology of liver and mandibular mass yielded a previously undescribed species of Schaalia, whose 16 S rRNA gene sequence was 97.0 % identical to Schaalia canis. Whole genome sequencing of the opossum isolate and calculation of average nucleotide identity confirmed a novel species of Schaalia, for which no whole genome sequence is yet available. CONCLUSIONS: The herewith reported Schaalia infection in the gray four-eyed opossum resembling classical actinomycosis gives a novel insight into new exotic animal bacterial diseases. Schaalia species may belong to the normal oral microbiome, as in macropods, and may serve as a contributor to opportunistic infections. Due to the lack of current literature, more insights and improved knowledge about Schaalia spp. and their pathogenicity will be useful to choose appropriate therapy regimens and improve the treatment success rate and outcome in exotic and endangered species.

Low occurrence of Bartonella in synanthropic mammals and associated ectoparasites in peri-urban areas from Central-Western and Southern Brazil.

Worldwide, Bartonella species are known to infect a wide range of mammalian and arthropod hosts, including humans. The current study aimed to investigate the prevalence of Bartonella spp. in synanthropic mammals captured in peri-urban areas from Central-Western and Southern Brazil and their ectoparasites. For this aim, 160 mammals belonging to four species, and 218 associated arthropods were sampled. DNA was extracted and subjected to different Bartonella screening assays. Additionally, blood samples from 48 small rodents were submitted to liquid BAPGM culture followed by qPCR assay and solid culture. Two out of 55 Rattus captured in Santa Catarina state were PCR-positive for Bartonella when targeting the nuoG, 16S, and ITS loci. Sequences showed high homology with Bartonella coopersplainsensis. Conversely, all 48 small rodents, 14 capybaras and 43 opossum DNA samples from animals trapped in Mato Grosso do Sul were Bartonella negative in the HRM real time PCR assays targeting the ITS locus and gltA gene. Additionally, all mammal-associated ectoparasites showed negativity results based on HRM real time PCR assays. The present study showed, for the first time, the occurrence of B. coopersplainsensis in Brazil, shedding some light on the distribution of rats-related Bartonella in South America. In addition, the majority of rodents and marsupials were negative for Bartonella spp. Since B. coopersplainsensis reservoirs - Rattus spp. - are widely dispersed around the globe, their zoonotic potential should be further investigated.

Detection of Hemotropic Mycoplasma sp. in white-eared opossums (Didelphis albiventris) from Southern Brazil / Detecção de Mycoplasma sp. hemotrópico em gambás-de-orelha-branca (Didelphis albiventris) no Sul do Brasil

Abstract Opossums are marsupials from the New World of the genus Didelphis and known as synanthropic animals due to their proximity with human beings. To date, 'Candidatus Mycoplasma haemodidelphis' has been solely found infecting the North American opossum (Didelphis virginiana). Accordingly, the aim of this study was to screen eight white-eared opossums (Didelphis albiventris) from a public park in Maringa city, Paraná State, southern Brazil, for hemoplasma infection. Blood samples were taken from caudal venipuncture, and DNA was extracted and further screened by a pan-hemoplasma PCR assay. Seven out of eight (87.50%; CI 95%: 47.35-99.68%) white-eared opossums were positive for Mycoplasma spp. Sequencing of the 16S rRNA fragment showed 98,97% identity with 'Ca. M. haemodidelphis' detected in the USA. Three out of eight (37.50%; CI 95%: 8.52-75.51%) white-eared opossums were infested by Amblyomma dubitatum ticks. This is the first report on detection of a potentially novel hemotropic Mycoplasma sp. infecting opossums from South America.

Resumo Gambás são marsupiais do Novo Mundo, pertencentes ao gênero Didelphis, e considerados animais sinantrópicos devido à sua proximidade com seres humanos. Atualmente, a espécie 'Candidatus Mycoplasma haemodidelphis' só foi encontrada infectando gambá norte americano (Didelphis virginiana). O objetivo do presente estudo foi detectar a infecção por hemoplasmas em oito gambás-de-orelha-branca (Didelphis albiventris) capturados em um parque público da cidade de Maringá, no Estado do Paraná, sul do Brasil. Amostras de sangue foram coletadas por venopunção caudal para a extração do DNA e posterior análise pela PCR para espécies de hemoplasmas. Sete de oito animais (87,50%; CI 95%: 47,35-99,68%) foram considerados positivos para Mycoplasma spp. O sequenciamento do fragmento do gene 16S rRNA obtido apresentou 98.97% de similaridade com sequências de 'Ca. M. haemodidelphis' detectadas nos Estados Unidos. Três gambás (37,50%; CI 95%: 8,52-75,51%) estavam infestados por carrapatos da espécie Amblyomma dubitatum. Esse é o primeiro relato de detecção de uma potencial nova espécie de Mycoplasma hemotrópico infectando gambás na América do Sul.

Detection of Hemotropic Mycoplasma sp. in white-eared opossums (Didelphis albiventris) from Southern Brazil.

Opossums are marsupials from the New World of the genus Didelphis and known as synanthropic animals due to their proximity with human beings. To date, 'Candidatus Mycoplasma haemodidelphis' has been solely found infecting the North American opossum (Didelphis virginiana). Accordingly, the aim of this study was to screen eight white-eared opossums (Didelphis albiventris) from a public park in Maringa city, Paraná State, southern Brazil, for hemoplasma infection. Blood samples were taken from caudal venipuncture, and DNA was extracted and further screened by a pan-hemoplasma PCR assay. Seven out of eight (87.50%; CI 95%: 47.35-99.68%) white-eared opossums were positive for Mycoplasma spp. Sequencing of the 16S rRNA fragment showed 98,97% identity with 'Ca. M. haemodidelphis' detected in the USA. Three out of eight (37.50%; CI 95%: 8.52-75.51%) white-eared opossums were infested by Amblyomma dubitatum ticks. This is the first report on detection of a potentially novel hemotropic Mycoplasma sp. infecting opossums from South America.

Salmonella, Campylobacter, Clostridium difficile, and anti-microbial resistant Escherichia coli in the faeces of sympatric meso-mammals in southern Ontario, Canada.

The role of free-ranging wildlife in the epidemiology of enteropathogens causing clinical illness in humans and domestic animals is unclear. Salmonella enterica and anti-microbial resistant bacteria have been detected in the faeces of raccoons (Procyon lotor), but little is known about the carriage of these bacteria in other sympatric meso-mammals. Our objectives were to: (a) report the prevalence of Salmonella and associated anti-microbial resistance, Campylobacter spp, Clostridium difficile, and anti-microbial resistant Escherichia coli in the faeces of striped skunks (Mephitis mephitis) and Virginia opossums (Didelphis virginiana) in southern Ontario; and (b) compare the prevalence of these bacteria in the faeces of these meso-mammal hosts with raccoons from a previously reported study. Faecal swabs were collected from striped skunks and Virginia opossums on five swine farms and five conservation areas from 2011 to 2013. Salmonella was detected in 41% (9/22) and 5% (5/95) of faecal swabs from Virginia opossums and striped skunks, respectively. None of the Salmonella serovars carried resistance to anti-microbials. The prevalence of Campylobacter spp., C. difficile, and anti-microbial resistant E. coli ranged from 6% to 22% in striped skunk and Virginia opossums. Using exact logistic regression, Salmonella was significantly more likely to be detected in faecal swabs of Virginia opossums than skunks and significantly less likely in faecal swabs from skunks than raccoons from a previously reported study. In addition, Campylobacter spp. was significantly more likely to be detected in raccoons than opossums. Salmonella Give was detected in 8/9 (89%) of Salmonella-positive Virginia opossum faecal swabs. Our results suggest that striped skunks and Virginia opossums have the potential to carry pathogenic enteric bacteria in their faeces. The high prevalence of Salmonella Give in Virginia opossum faecal swabs in this study as well as its common occurrence in other Virginia opossum studies throughout North America suggests Virginia opossums may be reservoirs of this serovar.

Pathogenic Leptospira species are widely disseminated among small mammals in Atlantic Forest biome.

Leptospirosis is a common worldwide bacterial zoonosis and has been studied in One Health approaches. Small mammals are described as the most important maintenance reservoirs of several pathogens in nature, including leptospires. The aim of this study was to identify infection by leptospires among small mammals on the Atlantic forest biome and evaluate their potential as carriers of these spirochetes. A total of 153 small mammals belonging to orders Rodentia and Didelphimorphia (distributed on 17 genera and 22 species) were captured. Blood and kidney samples were collected from animals and a conventional PCR targeted on lipL32 gene was conducted on renal tissues. Species identification was performed in eight samples by sequencing of rrs gene. A total of 28% of the animals presented lipL32 PCR-positive, and four pathogenic Leptospira species (L. interrogans, L. borgpetersenii, L. santarosai and L. noguchii) were identified. This study highlights the role of small mammals as carriers of leptospires on the Atlantic Forest representing a potential source of pathogenic Leptospira spp infection for both humans and domestic animals.

Newly Recognized Pediatric Cases of Typhus Group Rickettsiosis, Houston, Texas, USA.

An increase in typhus group rickettsiosis and an expanding geographic range occurred in Texas, USA, over a decade. Because this illness commonly affects children, we retrospectively examined medical records from 2008-2016 at a large Houston-area pediatric hospital and identified 36 cases. The earliest known cases were diagnosed in 2011.

Intrinsic and extrinsic factors related to pathogen infection in wild small mammals in intensive milk cattle and swine production systems.

BACKGROUND: Understanding the ecological processes that are involved in the transmission of zoonotic pathogens by small mammals may aid adequate and effective management measures. Few attempts have been made to analyze the ecological aspects that influence pathogen infection in small mammals in livestock production systems. We describe the infection of small mammals with Leptospira spp., Brucella spp., Trichinella spp. and Cysticercus fasciolaris and assess the related intrinsic and extrinsic factors in livestock production systems in central Argentina at the small mammal community, population and individual levels. METHODOLOGY/PRINCIPAL FINDINGS: Ten pig farms and eight dairy farms were studied by removal trapping of small mammals from 2008 to 2011. Each farm was sampled seasonally over the course of one year with cage and Sherman live traps. The 505 small mammals captured (14,359 trap-nights) included three introduced murine rodents, four native rodents and two opossums. Leptospira spp., anti-Brucella spp. antibodies and Trichinella spp. were found in the three murine rodents and both opossums. Rattus norvegicus was also infected with C. fasciolaris; Akodon azarae and Oligoryzomys flavescens with Leptospira spp.; anti-Brucella spp. antibodies were found in A. azarae. Two or more pathogens occurred simultaneously on 89% of the farms, and each pathogen was found on at least 50% of the farms. Pathogen infections increased with host abundance. Infection by Leptospira spp. also increased with precipitation and during warm seasons. The occurrence of anti-Brucella spp. antibodies was higher on dairy farms and during the winter and summer. The host abundances limit values, from which farms are expected to be free of the studied pathogens, are reported. CONCLUSIONS/SIGNIFICANCE: Murine rodents maintain pathogens within farms, whereas other native species are likely dispersing pathogens among farms. Hence, we recommend preventing and controlling murines in farm dwellings and isolating farms from their surroundings to avoid contact with other wild mammals.

Rickettsia amblyommii associado a roedores e marsupiais nativos da Estação Experimental Rafael Fernandes da UFERSA, Rio Grande do Norte / Rickettsia amblyommii associated with rodent and marsupials native of Raphael Fernandes Experimental Station da UFERSA, Rio Grande do Norte, Brazil

O presente estudo teve como objetivo registrar a ocorrência de Rickettsia sp. em roedores e marsupiais nativos da Estação Experimental Rafael Fernandes da UFERSA, Mossoró/RN. O trabalho consistiu em uma pesquisa de campo, com roedores e marsupiais silvestres, com os dados expressos em frequência simples e porcentagem através do programa estatístico IBM SPSS (Armonk, NY: IBM Corp.), versão 22.0. Coletaram-se amostras de plasma sanguíneo de marsupiais (36) e de roedores (5). Destes, 64 continham Amblyomma auricularium, 7 Amblyomma parvum e 12 Amblyomma sp. As amostras de plasma sanguíneo foram analisadas através da técnica de Reação de Imunofluorescência Indireta. Exemplares de A. auricularium e a A. parvum foram macerados e submetidos a Técnica de Reação em Cadeia da Polimerase. Das amostras de plasma testadas, 17,60% apresentaram soropositividade para Rickettsia amblyommii. Oito exemplares de A. auricularium estavam positivos para R. amblyommii na análise de fragmentos dos genes gltA (350 bp) e ompA (587 pb), com 100% de similaridade com Candidatus R. amblyommii estirpe Bahia e AaPE, corres­pondendo a uma baixa circulação do agente dentre os vetores e hospedeiros. Esta pesquisa registra pela primeira vez a ocorrência de R. amblyommii em marsupiais Gracilinanus agilis e Monodelphis domestica pertencentes a Família Didelphidae, e roedores das Famílias Echimyidae e Cricetidae, cujas espécies foram Thrichomys sp. e Wiedomys sp., respectivamente, em Mossoró, estado do Rio Grande do Norte.(AU)

The study aimed to register the occurrence of Rickettsia sp. in rodents and marsupials native of the Rafael Fernandes Experimental Station of UFERSA, Mossoró/RN, Brazil. The study consisted of field research on small wild mammals, with data expressed in simple frequency and percentage through IBM SPSS (IBM Corp., Armonk, NY), version 22.0. Samples of blood plasma from 36 marsupials and 5 rodents were collected. From these, 64 contained Amblyomma auricularium, 7 Amblyomma parvum and 12 Amblyomma sp. All blood plasma samples were analyzed by indirect immunofluorescence technique, and 16 macerated specimens of A. auricularium and 3 of A. parvum were analyzed by reaction technique Polymerase Chain. From the tested plasma samples 17.60% were seropositive for Rickettsia amblyommii, 8 were positive for A. auricularium e R. amblyommii in gene gltA analysis of the fragments (350 bp) and ompA (587 bp) with 100% similarity with Candidatus R. amblyommii Bahia and AAPE strain, what corresponded to a low circulation of the agent from the vectors and hosts. This study registers for the first time the occurrence of R. amblyommii in marsupials Gracilinanus agilis and Monodelphis domestica belonging to the Didelphidae family, and in rodents of the Echimyidae and Cricetidae families, the species of which were Thrichomys sp. and Wiedomys sp. respectively, in Mossoró, Rio Grande do Norte.(AU)

Posible transmisión oral de la enfermedad de Chagas en trabajadores del sector de los hidrocarburos en Casanare, Colombia, 2014 / Possible oral transmission of Chagas disease among hydrocarbons sector workers in Casanare, Colombia, 2014

RESUMEN Introducción. Trypanosoma cruzi, agente etiológico de la enfermedad de Chagas, puede transmitirse por vía oral tras la ingestión de alimentos o bebidas contaminadas. En la semana epidemiológica 14 del 2014, se notificaron dos casos de enfermedad aguda de Chagas en Paz de Ariporo, Casanare, entre trabajadores del sector de los hidrocarburos, episodio que motivó la investigación epidemiológica en el área. Objetivo. Caracterizar la población afectada, establecer medidas de control y confirmar el brote. Materiales y métodos. Se hizo una investigación de brote con los siguientes componentes: a) búsqueda de personas sintomáticas (cuadro clínico sugerente de enfermedad de Chagas según la definición de caso), para remitirlas a atención médica; b) aplicación de una encuesta entomológica en 192 de 197 viviendas; c) inspección sanitaria y análisis microbiológico de muestras de alimentos, y d) estudio de reservorios. La organización y el análisis de los datos se hicieron mediante estadística descriptiva con el programa Epi-Info 7.1.5. Asimismo, se establecieron los índices de infestación en el domicilio y el peridomicilio. Resultados. Se registraron 552 personas expuestas y se confirmaron por laboratorio 40 casos (7,2 %); siete casos se dieron en mujeres (17,5 %) y 33 en hombres (82,5%), es decir, en una relación de 1:5. La edad promedio fue de 39,1 (± 10,8) años, la tasa de ataque, de 7,2 %, y la letalidad, de 5 % (2/40). Los signos y síntomas incluyeron fiebre en el 100 % de los casos, cefalea en el 80 %, mialgias y artralgias en el 65 %, edema facial en el 55 %, y dolor abdominal en el 37,5 %. El tiempo promedio de incubación fue de 17 (3-21) días. El índice de infestación de Rhodnius prolixus fue de 3,3 % en el domicilio y de 2,2 % en el peridomicilio. En los cinco restaurantes inspeccionados, se encontraron condiciones sanitarias deficientes y alimentos con niveles de contaminación microbiológica inaceptables. Por último, un perro y dos zarigüeyas fueron positivos para los anticuerpos IgG en la prueba ELISA. Conclusiones. Mediante el análisis de las características epidemiológicas, ambientales y sanitarias, se confirmó un brote agudo de enfermedad de Chagas por exposición ocupacional y de posible transmisión oral, que podría ser el de mayor proporción reportado hasta la fecha en Colombia.

ABSTRACT Introduction: Trypanosoma cruzi, the etiological agent for Chagas disease, can be transmitted by oral intake of contaminated food or drinks. During epidemiological week 14 of 2014, two cases of acute Chagas disease were notified among hydrocarbons sector workers in Paz de Ariporo, Casanare. Objective: To characterize the affected population, to establish control and prevention measures and to confirm the outbreak. Materials and methods: We conducted an outbreak investigation that included the following components: a) Search for symptomatic people compatible with Chagas disease according to the case definition for their referral to medical services; b) entomological survey (192/197 houses); c) sanitary inspection and microbiological analysis of food samples; and d) study of reservoirs. Data management and analysis were done with Epi-Info 7.1.5 using descriptive statistics. We also calculated intradomicile and peridomicile triatomine infestation indexes. Results: We detected 552 exposed people; 40 had the disease (7.2%), of whom seven were women (17,5%) and 33, men (82.5%), i.e., a male-female ratio of 5:1. The mean age was 39.1 ± 10.8 years; the attack rate was 7.2% and lethality, 5% (2/40). Symptoms included fever (100% of cases), headache (80%), myalgia and arthralgia (65%), facial edema (55%), and abdominal pain (37.5%). The mean incubation time was 17 days (range: 3-21). Rhodnius prolixus domiciliary infestation index was 3.3 % and 2.2% in the peridomicile. In the five restaurants inspected sanitary conditions were deficient and food samples were microbiologically non-conforming. We found a dog and two opossums positive for IgG antibodies by ELISA. Conclusions: Environmental, sanitary and epidemiological conditions at the place confirmed an outbreak of Chagas diseases related to occupational exposure, possibly by oral transmission, which may be the largest to date in Colombia.

[Possible oral transmission of Chagas disease among hydrocarbons sector workers in Casanare, Colombia, 2014].

INTRODUCTION: Trypanosoma cruzi, the etiological agent for Chagas disease, can be transmitted by oral intake of contaminated food or drinks. During epidemiological week 14 of 2014, two cases of acute Chagas disease were notified among hydrocarbons sector workers in Paz de Ariporo, Casanare. OBJECTIVE: To characterize the affected population, to establish control and prevention measures and to confirm the outbreak. MATERIALS AND METHODS: We conducted an outbreak investigation that included the following components: a) Search for symptomatic people compatible with Chagas disease according to the case definition for their referral to medical services; b) entomological survey (192/197 houses); c) sanitary inspection and microbiological analysis of food samples; and d) study of reservoirs. Data management and analysis were done with Epi-Info 7.1.5 using descriptive statistics. We also calculated intradomicile and peridomicile triatomine infestation indexes. RESULTS: We detected 552 exposed people; 40 had the disease (7.2%), of whom seven were women (17,5%) and 33, men (82.5%), i.e., a male-female ratio of 5:1. The mean age was 39.1 ± 10.8 years; the attack rate was 7.2% and lethality, 5% (2/40). Symptoms included fever (100% of cases), headache (80%), myalgia and arthralgia (65%), facial edema (55%), and abdominal pain (37.5%). The mean incubation time was 17 days (range: 3-21). Rhodnius prolixus domiciliary infestation index was 3.3 % and 2.2% in the peridomicile. In the five restaurants inspected sanitary conditions were deficient and food samples were microbiologically non-conforming. We found a dog and two opossums positive for IgG antibodies by ELISA. CONCLUSIONS: Environmental, sanitary and epidemiological conditions at the place confirmed an outbreak of Chagas diseases related to occupational exposure, possibly by oral transmission, which may be the largest to date in Colombia.

Rickettsial Infections among Ctenocephalides felis and Host Animals during a Flea-Borne Rickettsioses Outbreak in Orange County, California.

Due to a resurgence of flea-borne rickettsioses in Orange County, California, we investigated the etiologies of rickettsial infections of Ctenocephalides felis, the predominant fleas species obtained from opossums (Didelphis virginiana) and domestic cats (Felis catus), collected from case exposure sites and other areas in Orange County. In addition, we assessed the prevalence of IgG antibodies against spotted fever group (SFGR) and typhus group (TGR) rickettsiae in opossum sera. Of the 597 flea specimens collected from opossums and cats, 37.2% tested positive for Rickettsia. PCR and sequencing of rickettsial genes obtained from C. felis flea DNA preparations revealed the presence of R. typhi (1.3%), R. felis (28.0%) and R. felis-like organisms (7.5%). Sera from opossums contained TGR-specific (40.84%), but not SFGR-specific antibodies. The detection of R. felis and R. typhi in the C. felis fleas in Orange County highlights the potential risk for human infection with either of these pathogens, and underscores the need for further investigations incorporating specimens from humans, animal hosts, and invertebrate vectors in endemic areas. Such studies will be essential for establishing a link in the ongoing flea-borne rickettsioses outbreaks.

[Mycobacterium bovis in wildlife of the dairy regions of Santa Fe (Argentina)]. / Mycobacterium bovis en fauna silvestre de la cuenca lechera de Santa Fe, Argentina.

Control eradication campaigns of bovine tuberculosis based on the «test and slaughter¼ approach were successful in many countries and regions; however, in some areas the infection persists and one of the main reasons is Mycobacterium bovis infection in wild life species. Argentina has applied the same approach since 1999, achieving progress in dairy cattle herds. Nonetheless, the wildlife role has never been investigated. The objective of this study was to determine if wildlife from the Santa Fe dairy area is infected with M. bovis. Wildlife species having a positive tuberculin skin test were captured in five dairy farms. Ninety five wildlife mammals were captured; M. bovis was recovered from 7 possums (Didelphys albiventris), from one fox (Lycolapex gimnocercus) and from one rat (Rattus norvegicus). None of the animals exhibited macroscopic lesions. The most frequently isolated M. bovis spoligotypes were types 34 (4 isolates) and 12 (3 isolates). Spoligotype 34 is the most frequently isolated type in Argentine cattle. The role of D. albiventris as spillover host of M. bovis is discussed in this study.

Rickettsial Infection in Animals, Humans and Ticks in Paulicéia, Brazil.

A previous study in Paulicéia Municipality, south-eastern Brazil, reported 9.7% of the Amblyomma triste ticks to be infected by Rickettsia parkeri, a bacterial pathogen that causes spotted fever in humans. These A. triste ticks were shown to be associated with marsh areas, where the marsh deer Blastocerus dichotomus is a primary host for this tick species. During 2008-2009, blood serum samples were collected from 140 horses, 41 dogs, 5 opossums (Didelphis albiventris) and 26 humans in farms from Pauliceia Municipality. Ticks were collected from these animals, from vegetation and from additional wildlife in these farms. Overall, 25% (35/140) of the horses, 7.3% (3/41) of the dogs, 3.8% (1/26) of the humans and 100% (5/5) of the opossums were seroreactive (titre ≥64) to spotted fever group (SFG) Rickettsia spp. Multivariate statistical analysis indicated that horses that were allowed to forage in the marsh were 4.8 times more likely to be seroreactive to spotted fever group (SFG) Rickettsia spp than horses that did not forage in the marsh. In addition, horses that had been living in the farm for more than 8.5 years were 2.8 times more likely to be seroreactive to SFG Rickettsia spp than horses that were living for ≤8.5 years. Ticks collected from domestic animals or from vegetation included Amblyomma cajennense, Amblyomma coelebs, Amblyomma dubitatum, Dermacentor nitens and Rhipicephalus microplus. By PCR analyses, only one pool of A. coelebs ticks from the vegetation was shown to be infected by rickettsiae, for which DNA sequencing revealed to be Rickettsia amblyommii. Ticks (not tested by PCR) collected from wildlife encompassed A. cajennense and Amblyomma rotundatum on lizards (Tupinambis sp), and A. cajennense and A. triste on the bird Laterallus viridis. Our results indicate that the marsh area of Paulicéia offers risks of infection by SFG rickettsiae.

Anaplasma phagocytophilum in small mammals and ticks in northeast Florida.

Human anaplasmosis is an emerging tick-borne disease in the United States, but few studies of the causative agent, Anaplasma phagocytophilum, have been conducted in southeastern states. The aim of this study was to determine if A. phagocytophilum is present in small mammals and ticks in northeast Florida. Polymerase chain reaction assays designed to amplify portions of the major surface protein 2 gene (p44), 16S rDNA, and groESL operons were used to test rodent blood and tick DNA samples for the presence of A. phagocytophilum. Positive samples were confirmed by DNA sequence analysis. Anaplasma phagocytophilum DNA was detected in less than 5% of cotton mice and 45% of cotton rats from two sites in northeast Florida. Anaplasma phagocytophilum DNA was also confirmed in 1.3% of host-seeking adult Ixodes scapularis tested and 2.7% of host-seeking adult Amblyomma americanum. This report describes the first DNA sequence data confirming strains of A. phagocytophilum in rodents and ticks in Florida. The DNA sequences of the msp2, 16S rDNA, and groESL gene fragments obtained in this study were highly similar to reference strains of human pathogenic strains of A. phagocytophilum. These findings suggest that A. phagocytophilum is present and established among some small mammal species in northeast Florida. Although the infection prevalence was low in the total number of ticks tested, the presence of A. phagocytophilum in two human biting tick species, one of which is a known competent vector, suggests that humans in this region may be at risk of granulocytic anaplasmosis caused by this pathogen.

Biodiversity of Clostridium botulinum type E associated with a large outbreak of botulism in wildlife from Lake Erie and Lake Ontario.

The genetic relatedness of Clostridium botulinum type E isolates associated with an outbreak of wildlife botulism was studied using random amplification of polymorphic DNA (RAPD). Specimens were collected from November 2000 to December 2008 during a large outbreak of botulism affecting birds and fish living in and around Lake Erie and Lake Ontario. In our present study, a total of 355 wildlife samples were tested for the presence of botulinum toxin and/or organisms. Type E botulinum toxin was detected in 110 samples from birds, 12 samples from fish, and 2 samples from mammals. Sediment samples from Lake Erie were also examined for the presence of C. botulinum. Fifteen of 17 sediment samples were positive for the presence of C. botulinum type E. Eighty-one C. botulinum isolates were obtained from plants, animals, and sediments; of these isolates, 44 C. botulinum isolates produced type E toxin, as determined by mouse bioassay, while the remaining 37 isolates were not toxic for mice. All toxin-producing isolates were typed by RAPD; that analysis showed 12 different RAPD types and multiple subtypes. Our study thus demonstrates that multiple genetically distinct strains of C. botulinum were involved in the present outbreak of wildlife botulism. We found that C. botulinum type E is present in the sediments of Lake Erie and that a large range of bird and fish species is affected.

Tick-borne disease agents in various wildlife from Mississippi.

Because tick-borne diseases are becoming increasingly important throughout the world, monitoring their causative agents in wildlife may serve as a useful indicator of potential human exposure. We assessed the presence of known and putative zoonotic, tick-borne agents in four wildlife species in Mississippi. Animals were tested for exposure to or infection with Ehrlichia chaffeensis, Ehrlichia ewingii, Borrelia lonestari, Rickettsia spp., Anaplasma phagocytophilum, and Francisella tularensis. Whole blood and serum were tested from white-tailed deer (WTD; Odocoileus virginianus) and feral swine (Sus scrofa); serum was tested from raccoons (Procyon lotor) and opossums (Didelphis virginiana). We used polymerase chain reaction to detect all agents in blood, whereas an indirect fluorescent antibody assay was used to detect antibodies to E. chaffeensis, B. lonestari, and Rickettsia parkeri (spotted fever group rickettsiae) antigens in serum. Molecular evidence of infection with E. chaffeensis, B. lonestari, and An. phagocytophilum was detected only in WTD. Antibodies to E. chaffeensis antigen were detected in 43.9% of WTD, 32.8% of swine, 42.1% of raccoons, and 15.8% of opossums. Serologic evidence of exposure to B. lonestari antigen was found in 19.3% of WTD, 6.9% of swine, and 5.3% of raccoons, but not in opossums. Interestingly, the percent of animals with antibodies reactive to spotted fever group rickettsiae (R. parkeri antigen) was highest in raccoons (73.7%) and opossums (57.9%). These results support the role of WTD as reservoirs for E. chaffeensis, B. lonestari, and An. phagocytophilum, as well as provide additional evidence for exposure of raccoons and opossums to E. chaffeensis. Finally, we provide new data that feral swine may have antibodies to these agents. Thus, in general, these four wildlife species are exposed to tick-borne disease agents in Mississippi, suggesting that ticks carry and have the potential to transmit the agents to humans in the state.

Epizootiologic survey of Mycobacterium bovis in wildlife and farm environments in northern Michigan.

Bovine tuberculosis (bovine TB), caused by Mycobacterium bovis, has reemerged in northern Michigan, USA, with detections in white-tailed deer (Odocoileus virginianus) in 1994 and in cattle in 1998. Since then, significant efforts have been directed toward reducing deer densities in the area in the hopes of reducing the bovine TB prevalence rate in deer and eliminating spillover of the disease into cattle. Despite the success of the efforts to reduce deer densities, additional cattle herds have become infected. Other mammals can be infected with M. bovis, and some carnivores and omnivores had been found to be infected with the disease in northern Michigan, USA. We conducted a multiyear surveillance effort to detect bovine TB in wild species of mammals in the Michigan, USA, outbreak area. From 2002 to 2004, tissue samples from 1,031 individual animals of 32 species were collected, processed, and cultured for M. bovis. Only 10 (1.0%) were culture-positive for M. bovis (five raccoons [Procyon lotor], four opossums [Didelphis virginiana], and one grey fox [Urocyon cinereoargenteus]). We also found two raccoons and four opossums to be positive for Mycobacterium avium. We collected 503 environmental samples from cattle farms recently identified as bovine TB positive; none yielded positive M. bovis culture results. Finally, we used infrared cameras to document wildlife use of four barns in the area. Many avian and mammalian species of wildlife were observed, with raccoons being the most commonly observed species. This surveillance study identified no new wildlife species that should be considered significant reservoirs of bovine TB in the outbreak area in northern Michigan, USA. However, the relatively high, apparent bovine TB prevalence rates in some carnivorous and omnivorous species, their relatively long life spans, and their frequent use of barns, suggests that removal of raccoons, opossums, foxes, and coyotes (Canis latrans) should be considered when a newly infected farm is depopulated of cattle.

Murine typhus in Austin, Texas, USA, 2008.

In August 2008, Texas authorities and the Centers for Disease Control and Prevention investigated reports of increased numbers of febrile rash illnesses in Austin to confirm the causative agent as Rickettsia typhi, to assess the outbreak magnitude and illness severity, and to identify potential animal reservoirs and peridomestic factors that may have contributed to disease emergence. Thirty-three human cases of confirmed murine typhus were identified. Illness onset was reported from March to October. No patients died, but 23 (70%) were hospitalized. The case-patients clustered geographically in central Austin; 12 (36%) resided in a single ZIP code area. Specimens from wildlife and domestic animals near case-patient homes were assessed; 18% of cats, 44% of dogs, and 71% of opossums had antibodies reactive to R. typhi. No evidence of R. typhi was detected in the whole blood, tissue, or arthropod specimens tested. These findings suggest that an R. typhi cycle involving opossums and domestic animals may be present in Austin.