A serological and molecular study on Francisella tularensis in rodents from Hamadan province,Western Iran

Introduction and purposeTularemia is a zoonotic disease, the most important hosts of which are rodents. Endemic regions and reservoirs of F. tularensis are not well-researched areas in Iran. The present study aimed to study F. tularensis infection in the rodent populations of western Iran.Materials and methodsSamples were collected in different areas of Kabudar Ahang County in Hamadan province (west of Iran) from 2014 to 2017. Tularemia serological and molecular tests were conducted using the tube agglutination test and Real-time PCR method tracking the ISFtu2 gene. Positive serum samples were evaluated for cross-reactivity with brucellosis.ResultsA total of 433 rodents, collected from 33 localities, were included in the study. The most abundant species belonged to the Persian jird (Meriones persicus; 75.5%), and Libyan jird (Meriones libycus; 10.1%). Among the studied samples, three (0.74 %) were seropositive and five (1.15%) were PCR positive. Seropositive samples were two M. persicus and one M. libycus, and PCR positive rodents were four M. persicus and one M. vinogradovi. Tularemia seropositive samples showed no cross-reactivity with brucellosis.ConclusionGiven the presence of infection in rodents with tularemia agent in the studied area, it is crucial to elucidate the risks of rodent exposure to tularemia for physicians, health personnel and the general population.     

Tularemia in Alaska, 1938 - 2010

Tularemia is a serious, potentially life threatening zoonotic disease. The causative agent, Francisella tularensis, is ubiquitous in the Northern hemisphere, including Alaska, where it was first isolated from a rabbit tick (Haemophysalis leporis-palustris) in 1938. Since then, F. tularensis has been isolated from wildlife and humans throughout the state. Serologic surveys have found measurable antibodies with prevalence ranging from < 1% to 50% and 4% to 18% for selected populations of wildlife species and humans, respectively. We reviewed and summarized known literature on tularemia surveillance in Alaska and summarized the epidemiological information on human cases reported to public health officials. Additionally, available F. tularensis isolates from Alaska were analyzed using canonical SNPs and a multi-locus variable-number tandem repeats (VNTR) analysis (MLVA) system. The results show that both F. t. tularensis and F. t. holarctica are present in Alaska and that subtype A.I, the most virulent type, is responsible for most recently reported human clinical cases in the state.     

Seroepidemiological study of Q fever,brucellosis and tularemia in butchers and slaughterhouses workers in Lorestan,western of Iran

Most zoonoses are occupational diseases. Q fever, brucellosis and tularemia are major zoonotic diseases for butchers and slaughterhouse workers. However, little information is available about these infectious diseases in such professional populations in western of Iran. The aim of this study was to investigate the seroprevalence and risk factors associated with these three zoonoses among butchers and slaughterhouse workers in the Lorestan province of Iran. In 2017, 289 individuals (144 butchers or slaughterhouse workers, and 145 people from the general population) were enrolled in 11 different counties of this province. Collected serum samples were tested by ELISA for detection of IgG antibodies against Coxiella burnetii, Brucella spp. or Francisella tularensis antigens. The seroprevalence of Q fever, brucellosis and tularemia among all participants were 23.5%, 31.8% and 3.8%, respectively. The seroprevalence of brucellosis and Q fever among butchers and slaughterhouse workers (43.7% and 29.8%, respectively) were significantly higher (p < 0.05) than those of the general population (20% and 17.2%, respectively). A contact history with small ruminants (sheep and goats) was associated with a higher risk of positive serology for all three studied zoonoses. The high seroprevalence for Q fever and brucellosis we found among butchers and slaughterhouse workers suggests that both diseases are common in these populations of the Lorestan province. Since these two infectious diseases are clinically unspecific, they must be systematically included in the etiological diagnosis of infectious diseases occurring in these at-risk populations. In addition, we recommend specific training programs as well as the use of personal protective equipment in these occupational groups to reduce the occurrence of these zoonotic diseases.     

Surveillance of Francisella tularensis in surface water of Kurdistan province,west of Iran

BackgroundThe etiologic agent of tularemia, Francisella tularensis, is transmitted to humans via ingestion of contaminated water or food, arthropods bite, respiratory aerosols, or direct contact with infected animals body fluids or tissues. In the current study, due to the importance of water in transmitting the disease and the report of the disease in different regions of Iran, surface water of Kurdistan province were evaluated for the presence of F.tularensis.Materials and MethodsSampling was carried out in five-counties of Kurdistan province. Sixty-six specimens of surface water were collected. The detection was carried out by targeting ISFtu2 and fopA genes using TaqMan real-time PCR. Moreover, the samples were both cultured and inoculated into NMRI inbreed mice. Spleens of inoculated mice and bacterial isolates were tested by TaqMan real-time PCR.ResultsDespite the lack of isolation of F. tularensis, the results of the molecular testing indicate the presence of bacteria in surface water. Molecular positivity of one sample (1.51%) was confirmed using a real-time PCR for both ISFtu2 and fopA genes. Moreover, 4.54% of the samples were positive for ISFtu2.ConclusionSince the in vitro isolation of bacteria from environmental samples is associated with a very low success rate and depends on various environmental parameters, the use of molecular techniques for monitoring of the bacteria in the contaminated areas is fully recommended.     

Tularemia

Tularemia is a potentially fatal multi-systemic disease of humans and other animals caused by the bacterial pathogen Francisella tularensis. The disease can be transmitted by ticks, biting flies, water exposure, food, and aerosols and occurs around the northern hemisphere including North America, Europe, and Asia. There are several defined species and subspecies, including F. tularensis subsp. tularensis (Jellison Type A) which is pathogenic for rabbits and occurs in North America, F. tularensis subsp. holarctica (Type B) and mediaasiatica which are less pathogenic for rabbits, and F. tularensis subsp. novicida which has been regarded sometimes as the separate species F. novicida. Because it can have a high aerosol-related infection rate, low infectious dose, and ability to induce fatal disease, F. tularensis is considered a potential agent of biological warfare and is classified by the US Department of Health and Human Services as a List A select agent. We discuss microbiological, clinicopathological, epidemiological, and ecological aspects of tularemia.     

An epidemiological survey on bovine and ovine babesiosis in Kurdistan Province, western Iran

The aim of this study was to determine the prevalence of the Babesia infection in domestic animals in Kurdistan Province of Iran for the first time. In this survey, 9,111 domestic livestock, including cattle and sheep, were randomly sampled and examined from 500 flocks in Kurdistan Province from July 2007 to September 2009. Thin peripheral blood smears were taken and then stained by Giemsa staining method. From a total of 9,111 collected samples, 2,642 were sheep and 6,469 were cattle. Babesia spp. is detected in 1,359 (51.4%) out of sheep samples and 136 (2.1%) out of cattle samples by direct examination of blood smear. Altogether, the prevalence rate of Babesia infection was 16.4% (n = 1,495) in both animal groups. Babesia ovis and Babesia bigemina were the most prevalent species found in sheep and cattle, respectively. The relatively high prevalence of Babesia infection in livestock indicates the epizootic stability status of babesiosis in the western part of Iran.     

Stakeholder Barriers and Benefits Associated With Improving Livestock Husbandry to Prevent Jaguar and Puma Depredation

Motivating ranchers to adopt preventive husbandry practices that limit livestock depredation by large carnivores, such as jaguars (Panthera onca) and pumas (Puma concolor), requires reducing perceived barriers and increasing benefits associated with coexistence. We assessed stakeholder perspectives on preventive practices by conducting eight focus groups consisting of ranchers, researchers, and government wildlife officers in Costa Rica using a nominal group technique to identify and rank benefits, barriers, and motivations. We identified 29 benefits, 27 positive motivations, 33 negative motivations, and 20 barriers. Common responses among stakeholders highlight the importance of economic issues, contextual factors, and external support. However, social interactions, a reactive approach to management, and personal motivations also influence rancher decision making, but tend to be ignored by researchers and wildlife officers. Nominal group rankings reveal misunderstandings and misalignment of priorities among stakeholders that should be targeted by collaborative problem-solving processes. Motivations behind prevention expose nuances of human–wildlife conflict.     

Detection of Leptospira spp. in wildlife reservoir hosts in Ontario through comparison of immunohistochemical and polymerase chain reaction genotyping methods

A total of 460 kidney samples from wildlife (beavers, coyotes, deer, foxes, opossums, otters, raccoons, skunks) were obtained from road-kill and hunter/trapper donations in Ontario between January 2010 and November 2012. The objectives of the study were to detect Leptospira spp. by immunohistochemistry and polymerase chain reaction (PCR), to map presence of leptospires in wildlife relative to livestock and human populations, and to characterize positive samples by sequencing and comparison to leptospires known to affect domestic animals and humans. The proportion of samples that tested positive ranged from 0% to 42%, with the highest rates in skunks and raccoons. Leptospira spp. were present in kidneys of wildlife across Ontario, particularly in areas of high human density, and areas in which livestock populations are abundant. The PCR was too weak in most samples to permit genotyping and examination of the relationship between the leptospires found in this study and those affecting domestic animals and humans.     

Tularaemia: A challenging zoonosis

In recent years, several emerging zoonotic vector-borne infections with potential impact on human health have been identified in Europe, including tularaemia, caused by Francisella tularensis. This remarkable pathogen, one of the most virulent microorganisms currently known, has been detected in increasingly new settings and in a wide range of wild species, including lagomorphs, rodents, carnivores, fish and invertebrate arthropods. Also, a renewed concern has arisen with regard to F. tularensis: its potential use by bioterrorists. Based on the information published concerning the latest outbreaks, the aim of this paper is to review the main features of the agent, its biology, immunology and epidemiology. Moreover, special focus will be given to zoonotic aspects of the disease, as tularaemia outbreaks in human populations have been frequently associated with disease in animals.     

Seroprevalence of Brucellosis,Tularemia, and Yersiniosis in Wild Boars (Sus scrofa) from North‐Eastern Germany

Brucellosis and tularemia are classical zoonotic diseases transmitted from an animal reservoir to humans. Both, wildlife and domestic animals, contribute to the spreading of these zoonoses. The surveillance of the animal health status is strictly regulated for domestic animals, whereas systematic disease monitoring in wildlife does not exist. The aim of the present study was to provide data on the prevalence of anti‐Brucella, anti‐Francisella and anti‐Yersinia antibodies in wild boars from North‐Eastern Germany to assess public health risks. A total of 763 sera of wild boars from Mecklenburg‐Western Pomerania hunted in 1995/1996 were tested using a commercially available Brucella suis ELISA, an in‐house lipopolysaccharide (LPS)‐based Francisella ELISA, and commercially available Western blot kits for the detection of anti‐Francisella and anti‐Yersinia antibodies. The Yersinia enterocolitica O:9 LPS is able to induce serological cross‐reactions indistinguishable from brucellosis due to a similar immunodominant epitope in the Brucella O‐polysaccharide. The Yersinia Western blot assay was, therefore, based on five recombinant Yersinia outer proteins which have been proved to be specific for the serodiagnosis of yersiniosis. Anti‐Brucella, anti‐Francisella and anti‐Yersinia antibodies were detected in 22.0%, 3.1%, and 62.6% of the wild boars, respectively. The high seroprevalence of tularemia and brucellosis in wild boars indicates that natural foci of these zoonoses are present in wildlife in Germany. However, the impact of transmission of zoonotic pathogens from wildlife to livestock is unknown. Only careful and systematic monitoring will help to prevent the (re)emergence of these zoonotic diseases in domestic animals and consequently human infection.     

Seroprevalence of tularemia in wild bears and hares in Japan

Tularemia is a zoonotic disease caused by Francisella tularensis. The distribution of the pathogen in Japan has not been studied well. In this study, seroprevalence of tularemia among wild black bears and hares in Japan was determined. Blood samples collected from 431 Japanese black bears (Ursus thibetanus japonicus) and 293 Japanese hares (Lepus brachurus) between 1998 and 2009 were examined for antibodies against F. tularensis by micro-agglutination test (MA) or enzyme-linked immunosorbent assay. By subsequent confirmatory tests using western blot (WB) and indirect immunofluorescence assay (IFA), eight sera from Japanese black bears were definitely shown to be seropositive. All of these eight bears were residents of the northeastern part of main-island of Japan, where human tularemia had been reported. On the other hand, no seropositive Japanese hares were found. These results suggest that Japanese black bears can serve as sentinel for tularemia surveillance and may help understand the distribution of F. tularensis throughout the country. This is the first report on detection of antibody to F. tularensis in black bears of Japan.     

Detection of contagious ovine ecthyma (orf) and risk factors for infection in small ruminants in Iran

The primary cause of contagious ecthyma is the orf virus, the parapoxvirus prototype. It is a viral problem observed in goat and sheep flocks in Iran, causing economic loss. Orf is a zoonosis with little epidemiological investigation present in Iran. The current research aims at determining the status of this virus, and a PCR was used as a confirmatory instrument. We sampled 668 goats and sheep and various breeding systems. Besides, the orf prevalence was studied, and vaccination efficacy was determined. Moreover, the potential risk factors surveyed for infection with ecthyma were identified. Samples were taken from goat and sheep flocks in the present cross-sectional research, and PCR was used for testing orf DNA. A checklist including animals’ general information was completed. Data were analyzed using univariate tests (chi-square and t-tests) and multivariable binary logistic regression analysis. Three hundred one (45%) goats and sheep detected orf DNA. The age of 70% of positive cases was below one month. Ecthyma infection was significantly higher in imported breeds (87.3%) than indigenous (39.3%). Ninety-six percent of infected goats and sheep in the present work were not vaccinated against ecthyma. The high prevalence of the orf virus was confirmed among goat and sheep flocks in Iran. It is necessary to train ranchers regarding sanitary actions, quarantine, and application of orf vaccination plans.     

Multi-species visit rates to farmyards: Implications for biosecurity

Disease transmission dynamics between wildlife and livestock are likely to be affected by contacts between the two populations. In this study, rates of visitation of rodents, badgers, red foxes and cats to 11 farmyards were quantified from July 2012 to August 2013. There was variation in rates of visitation between different species and between farmyards. Rates of visitation were 60.3% (n = 2374) for cats, 19.1% (n = 751) for rodents, 16.7% (n = 657) for red foxes and 3.9% (n = 153) for badgers. Multi-species visits are relevant to disease transmission, highlighting the need for increased biosecurity on farms.     

Influence of Ecosystem Services on Management Decisions by Public Land Ranchers in the Intermountain West,United States

Ecosystem services are benefits humans obtain as a result of ecosystem processes and conditions. In the western United States, public rangelands are managed for a spectrum of ecosystem services on behalf of multiple stakeholders. Decisions of ranchers who hold public land grazing allotments must balance operational needs for forage with societal expectations for other ecosystem services. To better understand their choices regarding ecosystem services, we interviewed ranchers to learn about the bases for their management decisions and identify services they believe rangelands provide. A total of 19 services were identified, many of which reflected ranchers’ recognition that they manage within the context of a broader social-ecological system (e.g., maintaining open space). We then conducted a mail survey of Bureau of Land Management grazing permittees in six states to understand the importance they personally place on different services, as well as the extent to which they manage with those services in mind. Fourteen of the 19 ecosystem services identified in interviews were reported by at least 50% of the survey sample (N = 435) as influencing their management decisions. Most respondents reported trying to manage deeded and leased land to the same standard. Aside from forage for livestock (ranked #1), ranchers were less likely to report managing for provisioning services than for cultural, supporting, and regulating services. Importance ratings for ecosystem services followed a similar pattern, although there were a few differences in rank order. Ranchers tended to report managing for more ecosystem services if they had larger operations, earned at least 50% of their income from ranching, spent more time out on the ranch, and relied on multiple sources for information about range management. Results indicate public land ranchers believe they are managing for multifunctionality, balancing their own operational needs with those of society.     

Seroprevalence of Toxoplasma gondii and Neospora caninum in sheep intended for human consumption in the Rondônia state,Western Brazilian Amazon

The aim of this study was to carry out a seroepidemiological study of anti-T. gondii and anti-N. caninum antibodies in serum samples from sheep intended for human consumption in the Rondônia state, in the Western Brazilian Amazon, and identify possible risk factors associated with seropositivity. The analysis of the 616 sheep serum samples revealed animal-level seroprevalence of T. gondii in the order of 52.4 % and of N. caninum of 60.6 %. Prevalence of co-infections was 33.4 %. The herd-level seroprevalence was 100 % for both the parasites. Applying of multivariate logistic regression analysis, test of the association between seroprevalence of T. gondii infection and the potential predictors showed that feed, fetal malformation, abortion, neurological problems (lambs), presence of rodents were the significant (P < 0.05) predictors. The variables presence of rodents, cats, wild animals and main activity showed statistical association (P < 0.05) with the seroprevalence of N. caninum. Due to the high prevalence of T. gondii and N. caninum found in this study, the absence of specific slaughterhouses for sheep and raw or undercooked sheep meat for human consumption we concluded that sheep can represent an important source of infection for humans.     

Role of Chlamydophila abortus in ovine and caprine abortion in Sardinia,Italy

Between 1999–2003, 14 321 sera and 646 abortion samples (498 foetuses and 148 placentae) were analysed from 807 sheep and goat farms distributed all over the island of Sardinia. After notification of abortion in a flock, sera collected at random from adult animals were examined to detect antibodies specific to Chlamydophila (C.) abortus by ELISA, whereas foetuses and placenta were analysed by PCR assay. Specific IgG antibodies were detected in 611 (4.8%) sheep and 106 (5.8%) goats. From a total of 2050 ovine and 151 caprine fetal samples including muscle, liver, abomasum, spleen, brain and placenta, 29 (1.4%) ovine and 1 (0.6%) caprine samples were C. abortus PCR-positive. Placenta was the tissue with the highest detection rate. These results indicate that the seroprevalence of C. abortus infection in sheep and goats is very low in Sardinia, and PCR results demonstrate that C. abortus has no significant role in abortion, especially in goats.     

Explaining Cattle Rancher Participation in Wildlife Conservation Technical Assistance Programs in the Southeastern United States

US natural resources and wildlife agencies have been increasing their efforts to involve cattle ranchers in wildlife conservation through technical assistance programs that provide for wildlife conservation activities. Understanding why ranchers choose to be involved in these programs is fundamental to increasing participation and ensuring their success. Using the theory of planned behavior as a theoretical model, we surveyed 1 093 ranchers in Alabama, Florida, Georgia, and Mississippi to explain and predict intention to participate in technical assistance programs, specifically, wildlife workshops and field days. All three theory components—attitude, subjective norm, and perceived behavioral control—were important to intent to participate and explained 41% of the variance, with perceived behavioral control and subjective norm having the greatest standardized effects (β = 0.329 and β = 0.316, respectively). Investigation of the construct components yielded insight into how agencies could increase participation. Ranchers generally held positive attitudes toward wildlife workshops, perceiving them to be a good way to learn about wildlife management and perceiving that most ranches were suitable for wildlife, an instance of perceived behavioral control. However, ranchers did not perceive that workshops and field days were widely advertised or promoted, limiting the amount of perceived control they had over their participation. Additionally, ranchers identified normative groups whose opinions were important to them, namely their families, friends and neighbors, fellow ranchers, and agency staff. However, these same groups were not seen to actively encourage ranchers to participate in technical field days and workshops. Using key members of these normative groups to advertise and promote workshops and field days among their peers should increase rancher behavioral control and attitudes associated with technical workshops and field days. Employing strategies from this research to increase attendance at technical workshops and field days should improve wildlife conservation technical assistance program effects.     

Complex epidemiology and zoonotic potential for Cryptosporidium suis in rural Madagascar

Cryptosporidium spp. is the most important parasitic diarrheal agent in the world, is among the top four causes of moderate-to-severe diarrheal disease in young children in developing nations, and is problematic as an opportunistic co-infection with HIV. In addition, Cryptosporidium is a persistent challenge for livestock production. Despite its zoonotic potential, few studies have examined the ecology and epidemiology of this pathogen in rural systems characterized by high rates of overlap among humans, domesticated animals, and wildlife. To improve our understanding of the zoonotic potential of Cryptosporidium species in the rural tropics, we screened humans, livestock, peridomestic rodents, and wildlife using PCR-RFLP and sequencing-based approaches to distinguish species of Cryptosporidium in rural southeastern Madagascar. Cryptosporidium of multiple species/genotypes were apparent in this study system. Interestingly, C. suis was the dominant species of Cryptosporidium in the region, infecting humans (n = 1), cattle (n =  18), pigs (n= 3), and rodents (n = 1). The broad species range of C. suis and the lack of common cattle Cryptosporidium species (Cryptosporidium parvum and Cryptosporidium andersoni) in this system are unique. This report represents the fifth confirmed case of C. suis infection in humans, and the first case in Africa. Few rural human and livestock populations have been screened for Cryptosporidium using genus-specific genotyping methods. Consequently, C. suis may be more widespread in human and cattle populations than previously believed.     

The transmission of Cowdria ruminantium by Amblyomma sparsum

Transmission of Cowdria ruminantium (heartwater) by nymphs of Amblyomma sparsum has been demonstrated. Adults of the same tick species failed to transmit the disease after feeding on infected sheep in either the larval or the nymphal stage, and there was no transovarian transmission. A. sparsum seldom parasitizes domestic livestock but could be of importance in maintaining reservoirs of heartwater infection in wildlife populations.     

Molecular and Statistical Analysis of Campylobacter spp. and Antimicrobial‐Resistant Campylobacter Carriage in Wildlife and Livestock from Ontario Farms

The objectives of this study were to (i) compare the carriage of Campylobacter and antimicrobial‐resistant Campylobacter among livestock and mammalian wildlife on Ontario farms, and (ii) investigate the potential sharing of Campylobacter subtypes between livestock and wildlife. Using data collected from a cross‐sectional study of 25 farms in 2010, we assessed associations, using mixed logistic regression models, between Campylobacter and antimicrobial‐resistant Campylobacter carriage and the following explanatory variables: animal species (beef, dairy, swine, raccoon, other), farm type (swine, beef, dairy), type of sample (livestock or wildlife) and Campylobacter species (jejuni, coli, other). Models included a random effect to account for clustering by farm where samples were collected. Samples were subtyped using a Campylobacter‐specific 40 gene comparative fingerprinting assay. A total of 92 livestock and 107 wildlife faecal samples were collected, and 72% and 27% tested positive for Campylobacter, respectively. Pooled faecal samples from livestock were significantly more likely to test positive for Campylobacter than wildlife samples. Relative to dairy cattle, pig samples were at significantly increased odds of testing positive for Campylobacter. The odds of isolating Campylobacter jejuni from beef cattle samples were significantly greater compared to dairy cattle and raccoon samples. Fifty unique subtypes of Campylobacter were identified, and only one subtype was found in both wildlife and livestock samples. Livestock Campylobacter isolates were significantly more likely to exhibit antimicrobial resistance (AMR) compared to wildlife Campylobacter isolates. Campylobacter jejuni was more likely to exhibit AMR when compared to C. coli. However, C. jejuni isolates were only resistant to tetracycline, and C.  coli isolates exhibited multidrug resistance patterns. Based on differences in prevalence of Campylobacter spp. and resistant Campylobacter between livestock and wildlife samples, and the lack of similarity in molecular subtypes and AMR patterns, we concluded that the sharing of Campylobacter species between livestock and mammalian wildlife was uncommon.