Major Reservoir Hosts Research Articles

First record of Leishmaniasis in wild Jaguars (Panthera onca) from Mexico

Leishmaniasis is a vector-borne disease and parasitic zoonosis that occurs worldwide (i.e., Latin America, Africa and Eurasian countries; Desjeux 2005, Dahroug et al. 2010, 2011, Otranto et al. 2013). It is caused by parasitic kinetoplastids of the Leishmania genus transmitted by arthropod vectors (i.e. sandflies such as Lutzomyia spp.) and presents a great diversity of clinical profiles, depending on the affected areas (Otranto et al. 2013). This disease mainly affects humans and wild carnivoran species, mostly canids, with dogs as major reservoir hosts (Deane & Deane 1954, Silva et al. 2000, Figueiredo et al. 2008, Dantas-Torres et al. 2012). Even though domestic and wild felids are unusual reservoirs, and rarely develop the disease, several studies suggest that these species may play an important role in the epidemiology of the disease (Poli et al. 2002, Maia et al. 2008, Nasereddin et al. 2008, Dantas-Torres et al. 2012). Here we report the first record of a clinical manifestation of Leishmaniasis disease in a wild Jaguar (Panthera onca) in Quintana Roo, Mexico.

Avian influenza virus exhibits distinct evolutionary dynamics in wild birds and poultry.

BackgroundWild birds are the major reservoir hosts for influenza A viruses, occasionally transmitting to other species such as domesticated poultry. Despite an abundance of genomic data from avian influenza virus (AIV), little is known about whether AIV evolves differently in wild birds and poultry, although this is critical to revealing the dynamics and time-scale of viral evolution. In particular, because environmental (water-borne) transmission is more common in wild birds, which may reduce the number of replications per unit time, it is possible that evolutionary rates are systematically lower in wild birds than in poultry.ResultsWe estimated rates of nucleotide substitution in two AIV subtypes that are strongly associated with infections in wild birds – H4 and H6 – and compared these to rates in the H5N1 subtype that has circulated in poultry for almost two decades. Our analyses of three internal genes confirm that H4 and H6 viruses are evolving significantly more slowly than H5N1 viruses, suggesting that evolutionary rates of AIV are reduced in wild birds. This result was verified by the analysis of a poultry-associated H6 lineage that exhibited a markedly higher substitution rate than those H6 viruses circulating in wild birds. Interestingly, we also observed a significant difference in evolutionary rate between H4 and H6, despite frequent reassortment rate among them.ConclusionsAIV experiences markedly different evolutionary dynamics between wild birds and poultry. These results suggest that rate heterogeneity among viral subtypes and ecological groupings should be taken into account when estimating evolutionary rates and divergence times.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-015-0410-5) contains supplementary material, which is available to authorized users.

Water buffalo as sentinel animals for schistosomiasis surveillance.

Introduction About 75% of human pathogens are zoonotic, meaning that they are communicated from animals to humans. (1) Schistosomiasis is a zoonotic disease with a complex transmission cycle involving aquatic snails and least 40 species of mammals, which serve as reservoir hosts. Worldwide, more than 700 million people are at risk of schistosomiasis and over 240 million people are infected with the parasite. (2) Schistosomiasis has been eliminated in Japan and the coastal plains of China by a combination of medical treatment, health education, improved water quality and sanitation and snail control through environmental modification, molluscicide and new farming methods. The World Health Organization (WHO) targets elimination of schistosomiasis in the Eastern Mediterranean, the Caribbean, Indonesia and the Mekong river basin by 2015, and in the Western Pacific, the Americas and selected African countries by 2020. (3) Here we argue that elimination guidelines for schistosomiasis should include surveillance of the animal reservoir. Transmission patterns Mapping of the transmission patterns in humans and animals can lead to a better understanding of transmission of schistosomiasis between different host species. Parts of China, Indonesia and the Philippines are endemic for schistosomiasis caused by Schistosoma japonicum. Cattle, water buffalo, goats, dogs, pigs and rats are potentially important reservoir hosts for this parasite because of their contact with humans. Other animals including cats and horses are not significant contributors because of their limited contact with contaminated water. (4) Grazing ruminants including goats, cattle and water buffalo are exposed to the parasite in transmission sites such as rice paddies. However, goats do not contribute much to transmission as they are uncommon in areas where schistosomiasis is endemic. Cattle and water buffalo are the major animal reservoirs contributing to human transmission of S. japonicum. (4-7) In China, bovine defecation is thought to be the main cause of environmental contamination. (6) Molecular studies have also suggested that transmission is through bovines rather than other domesticated animals. Cattle are less capable of recovering from infection than water buffalo. (8) Despite the fact that buffalo are capable of clearing the parasite, they are a major reservoir host because they are repeatedly exposed to the parasite when pulling ploughs in rice paddies. (9) In rice-growing regions, water buffalo are a good choice of sentinel animal. In Anhui and Sichuan provinces of China, re-emergence of schistosomiasis in humans was attributed to the high prevalence of schistosome infection among cows and water buffalo. (10,11) These human infections could be avoided by setting up a sensitive animal surveillance system to detect infection in animals and, by treating them promptly, preventing further contamination of the environment. Diagnostic tests Microscopic examination of stool samples is a standard test for several parasitic diseases, including schistosomiasis. However, microscopy has low sensitivity in diagnosing schistosomiasis among water buffalo due to the large volume of bovine excreta. Therefore, a more sensitive and specific diagnostic test is needed to identify schistosomiasis-free areas and prevent re-emergence of the disease. In previous studies, polymerase chain reaction (PCR) (12) and recombinant antigen-based enzyme-linked immunosorbent assay (ELISA) (13) have been found to be useful in identifying S. japonicum in stool samples. Surveillance can also be carried out in abattoirs where cattle and buffalo are slaughtered. Documentation of the original source of livestock then allows schistosome-infested sites to be located. Medical and veterinary approaches The complexity of zoonotic schistosomiasis transmission has been a major hindrance to elimination of the disease. …

Broad diversity of host responses of the white-footed mouse Peromyscus leucopus to Borrelia infection and antigens

Peromyscus leucopus, the white-footed mouse, is one of the more abundant mammals of North America and is a major reservoir host for at least five tickborne diseases of humans, including Lyme disease and a newly-recognized form of relapsing fever. In comparison to Mus musculus, which is not a natural reservoir for any of these infections, there has been little research on experimental infections in P. leucopus. With the aim of further characterizing the diversity of phenotypes of host responses, we studied a selection of quantitative traits in colony-bred and -reared outbred P. leucopus adults that were uninfected, infected with the relapsing fever agent Borrelia hermsii alone, or infected after immunization with Lyme disease vaccine antigen OspA and keyhole limpet hemocyanin (KLH). The methods included measurements of organ weights, hematocrits, and bleeding times, quantitative PCR for bacterial burdens, and enzyme immunoassays for serum antibodies against both the immunization proteins and cellular antigens of the infecting organism. The results included the following: (i) uninfected animals displayed wide variation in relative sizes of their spleens and in their bleeding times. (ii) In an experiment with matched littermates, no differences were observed between females and males at 7 days of infection in bacterial burdens in blood and spleen, relative spleen size, or antibody responses to the B. hermsii specific-antigen, FbpC. (iii) In studies of larger groups of males or females, the wide variations between bacterial burdens and in relative spleen sizes between individuals was confirmed. (iv) In these separate groups of males and females, all animals showed moderate-to-high levels of antibodies to KLH but wide variation in antibody levels to OspA and to FbpC. The study demonstrated the diversity of host responses to infection and immunization in this species and identified quantitative traits that may be suitable for forward genetics approaches to reservoir-pathogen interactions.

High prevalence of Schistosoma japonicum and Fasciola gigantica in bovines from Northern Samar, the Philippines.

The cause of zoonotic schistosomiasis in the Philippines is Schistosoma japonicum, which infects up to 46 mammalian hosts, including humans and bovines. In China, water buffaloes have been identified as major reservoir hosts for schistosomiasis japonica, contributing up to 75% of human transmission. In the Philippines, water buffaloes (carabao; Bubalus bubalis carabanesis) have, historically, been considered unimportant reservoirs. We therefore revisited the possible role of bovines in schistosome transmission in the Philippines, using the recently described formalin-ethyl acetate sedimentation (FEA-SD) technique and a qPCR assay to examine fecal samples from 153 bovines (both carabao and cattle) from six barangays in Northern Samar. A high prevalence of S. japonicum was found using qPCR and FEA-SD in both cattle (87.50% and 77.08%, respectively) and carabao (80.00% and 55.24%, respectively). The average daily egg output for each bovine was calculated at 195,000. High prevalence and infection intensity of F. gigantica was also found in the bovines by qPCR and FEA-SD (95.33% and 96.00%, respectively). The identification of bovines as major reservoir hosts for S. japonicum transmission suggests that bovine treatment and/or vaccination, as one becomes available, should be included in any future control program that aims to reduce the disease burden due to schistosomiasis in the Philippines.

Vaccines and diagnostics for zoonotic schistosomiasis japonica.

Schistosomiasis is one of the most prevalent, insidious and serious of the tropical parasitic diseases. Although the effective anthelmintic drug, praziquantel, is widely available and cheap, it does not protect against re-infection, drug-resistant schistosome may evolve and mass drug administration programmes based around praziquantel are probably unsustainable long term. Whereas protective anti-schistosome vaccines are not yet available, the zoonotic nature of Schistosoma japonicum provides a novel approach for developing a transmission-blocking veterinary vaccine in domestic animals, especially bovines, which are major reservoir hosts, being responsible for up to 90% of environmental egg contamination in China and the Philippines. However, a greater knowledge of schistosome immunology is required to understand the processes associated with anti-schistosome protective immunity and to reinforce the rationale for vaccine development against schistosomiasis japonica. Importantly as well, improved diagnostic tests, with high specificity and sensitivity, which are simple, rapid and able to diagnose light S. japonicum infections, are required to determine the extent of transmission interruption and the complete elimination of schistosomiasis following control efforts. This article discusses aspects of the host immune response in schistosomiasis, the current status of vaccine development against S. japonicum and reviews approaches for diagnosing and detecting schistosome infections in mammalian hosts.

High levels of Trypanosoma cruzi DNA determined by qPCR and infectiousness to Triatoma infestans support dogs and cats are major sources of parasites for domestic transmission

The competence of reservoir hosts of vector-borne pathogens is directly linked to its capacity to infect the vector. Domestic dogs and cats are major domestic reservoir hosts of Trypanosoma cruzi, and exhibit a much higher infectiousness to triatomines than seropositive humans. We quantified the concentration of T. cruzi DNA in the peripheral blood of naturally-infected dogs and cats (a surrogate of intensity of parasitemia), and evaluated its association with infectiousness to the vector in a high-risk area of the Argentinean Chaco. To measure infectiousness, 44 infected dogs and 15 infected cats were each exposed to xenodiagnosis with 10–20 uninfected, laboratory-reared Triatoma infestans that blood-fed to repletion and were later individually examined for infection by optical microscopy. Parasite DNA concentration (expressed as equivalent amounts of parasite DNA per mL, Pe/mL) was estimated by real-time PCR amplification of the nuclear satellite DNA. Infectiousness increased steeply with parasite DNA concentration both in dogs and cats. Neither the median parasite load nor the mean infectiousness differed significantly between dogs (8.1Pe/mL and 48%) and cats (9.7Pe/mL and 44%), respectively. The infectiousness of dogs was positively and significantly associated with parasite load and an index of the host’s body condition, but not with dog’s age, parasite discrete typing unit and exposure to infected bugs in a random-effects multiple logistic regression model. Real-time PCR was more sensitive and less time-consuming than xenodiagnosis, and in conjunction with the body condition index, may be used to identify highly infectious hosts and implement novel control strategies.

Endemicity ofOpisthorchis viverriniLiver Flukes, Vietnam, 2011–2012

Endemicity of<i>Opisthorchis viverrini</i>Liver Flukes, Vietnam, 2011–2012

Leptospirosis in the western Indian Ocean islands: what is known so far?

In the past decade, leptospirosis has emerged as a major zoonosis with a worldwide distribution. The disease is caused by bacteria of the genus Leptospira. The western Indian Ocean includes more than one hundred tropical or subequatorial islands where leptospirosis constitutes a major public health problem. The clinical signs of the human disease are generally similar to an influenza-like syndrome, but acute forms of the disease are reported and mortality remains significant in this region. In animals, clinical forms are mainly asymptomatic but leptospirosis reduces the fertility of livestock, resulting in economic losses. The data available about human and animal leptospirosis in the western Indian Ocean islands are diverse: human leptospirosis has been extensively studied in Reunion Island, Mayotte, and the Seychelles, whereas the human clinical disease has never been described in Madagascar, Comoros, Mauritius, or Rodrigues, mainly because of the deficiency in appropriate medical and diagnostic structures. The rat is recognized as the major reservoir host for the bacteria on all islands, but recent data from Reunion Island indicates that almost all mammals can be a source of contamination. The incidence of leptospirosis in humans is highly seasonal, and linked to the rainy season, which is favorable for the environmental maintenance and transmission of the bacteria. The epidemiology of leptospirosis is fully island-dependent, related to the number of mammalian species, the origins of the introduced mammalian species, the relationships between humans and fauna, and environmental as well as cultural and socio-economic factors.

Detection of Trypanosoma cruzi infection in naturally infected dogs and cats using serological, parasitological and molecular methods

Domestic dogs and cats are major domestic reservoir hosts of Trypanosoma cruzi and a risk factor for parasite transmission. In this study we assessed the relative performance of a polymerase chain reaction assay targeted to minicircle DNA (kDNA-PCR) in reference to conventional serological tests, a rapid dipstick test and xenodiagnosis to detect T. cruzi infection in dogs and cats from an endemic rural area in northeastern Argentina. A total of 43 dogs and 13 cats seropositive for T. cruzi by an immunosorbent assay (ELISA) and an indirect hemagglutination assay (IHA), which had been examined by xenodiagnosis, were also tested by kDNA-PCR. kDNA-PCR was nearly as sensitive as xenodiagnosis for detecting T. cruzi-infectious dogs and cats. kDNA-PCR was slightly more sensitive than xenodiagnosis in seropositive dogs (91% versus 86%, respectively) and cats (77% against 54%, respectively), but failed to detect all of the seropositive individuals. ELISA and IHA detected all xenodiagnosis-positive dogs and both outcomes largely agreed (kappa coefficient, κ=0.92), whereas both assays failed to detect all of the xenodiagnosis-positive cats and their agreement was moderate (κ=0.68). In dogs, the sensitivity of the dipstick test was 95% and agreed closely with the outcome of conventional serological tests (κ=0.82). The high sensitivity of kDNA-PCR to detect T. cruzi infections in naturally infected dogs and cats supports its application as a diagnostic tool complementary to serology and may replace the use of xenodiagnosis or hemoculture.

Serological Survey of Dogs From Egypt for Antibodies toLeishmaniaSpecies

Leishmaniasis is an insect-transmitted parasitic disease with a worldwide distribution. Leishmania spp. infections cause a broad spectrum of clinical signs, ranging from skin lesions to fatal visceral disease. Dogs are a major reservoir host for visceral leishmaniasis in humans. While the disease is endemic in the Middle East and North Africa, little is known concerning canine Leishmania spp. infections in Egypt. Accordingly, blood samples were collected from 50 stray dogs in Giza, Egypt. Canine sera were tested for antibodies to visceralizing Leishmania spp. by commercial immunochromatographic strip assays based on recombinant antigen K39. Antibodies to Leishmania spp. were found in 5 of 50 (10%) of dogs tested from Egypt. Results from this study indicate that stray dogs are exposed to visceralizing Leishmania species in Egypt.

Detection of human filarial parasite Brugia malayi in dogs by histochemical staining and molecular techniques

Human filariasis caused by Brugia malayi is still a public health problem in many countries of Asia including India, Indonesia, Malaysia and Thailand. The World Health Organization (WHO) has targeted to eliminate filariasis by the year 2020 by Mass annual single dose Diethylcarbamazine Administration (MDA). Results of the MDA programme after the first phase was less satisfactory than expected. Malayan filariasis caused by B. malayi is endemic in the south of Thailand where domestic cat serves as the major reservoir host. There is no report about the occurrence of B. malayi in dogs. The present work was carried out to find out the incidence of microfilariasis in dogs and also to detect the presence of human filarial infection in dogs, if any.One hundred dogs above 6 months of age presented to the veterinary college Hospital, Mannuthy, Kerala, with clinical signs suggestive of microfilariasis – fever, anorexia, conjunctivitis, limb and scrotal oedema – were screened for microfilariae by wet film examination. Positive cases were subjected to Giemsa staining, histochemical staining and molecular techniques. Results of the study showed that 80% of dogs had microfilariasis; out of which 20% had sheathed microfilaria. Giemsa and histochemical staining character, PCR and sequencing confirmed it as B. malayi. High prevalence of B. malayi in dogs in this study emphasized the possible role of dogs in transmission of human filariasis.

Identification of candidate host proteins that interact with LipL32, the major outer membrane protein of pathogenic Leptospira, by random phage display peptide library

Leptospirosis is a worldwide zoonotic disease caused by pathogenic Leptospira spp. Rodent species are the major reservoir hosts that can excrete leptospires in their urine leading to environmental contamination. After gaining entry into the host via skin breaks and mucosa, leptospires disseminate through the bloodstream to target organs causing a wide range of disease manifestations in susceptible mammalian hosts. The crucial step of infection requires host–pathogen interactions. LipL32, the major outer membrane protein (OMP) of pathogenic Leptospira, is conserved among pathogenic leptospires, immunogenic, and expressed in target organs during acute infection in animal models. Therefore, it may play a key role in host–microbe interactions. To identify host proteins that interact with LipL32, phage display technology was employed in our study. Recombinant LipL32 was used as a target molecule for biopanning with a random heptapeptide phage library to enrich for phages expressing peptides with high affinity to LipL32. After three rounds of panning, 44 plaques of eluted phages were subjected to pyrosequencing. Six different peptide sequences were identified and used to search for matching proteins in the database. Putative proteins with potential binding to LipL32 are proteins known to be expressed on the surface of target cells of pathogenic Leptospira such as chloride channel accessory 2, glycoprotein VI, scavenger receptor expressed by endothelial cell isoform I (SREC-I), coronin 2A, laminin alpha 5, collagen XX, and prostaglandin receptor EP1. However, interactions of LipL32 with these host proteins and their role in the pathogenesis of leptospirosis requires experimental confirmation.

Linking disease and community ecology through behavioural indicators: immunochallenge of white-footed mice and its ecological impacts.

1. Pathogens and immune challenges can induce changes in host phenotype in ways that indirectly impact important community interactions, including those that affect host-pathogen interactions. 2. To explore host behavioural response to immune challenge, we exposed wild white-footed mice (Peromyscus leucopus) to an immunogen from an endemic, zoonotic pathogen, the spirochete Borrelia burgdorferi. White-footed mice are a major reservoir host of Lyme disease (LD) spirochetes in northeastern USA and an abundant member of forest communities. The activity patterns, foraging behaviour, and space use of white-footed mice have implications for population growth rates of community members upon which mice incidentally prey (i.e. gypsy moths and native thrushes), as well as potentially determining host-vector encounter rates and human risk of LD. 3. Immunochallenge led to specific humoral (antibody) and cellular (i.e. elevated neutrophils and eosinophils) immune responses, supporting use of the immunogen as a surrogate for pathogenic infection. 4. Immunochallenged mice had reduced wheel-running activity early in the night when measured in the lab. However, mouse activity, as measured by track plates in natural field experiments, did not differ between mice exposed to the immunogen and unexposed mice. 5. Foraging behaviour of wild mice in the field - assessed with giving-up densities of seed at artificial feeding stations - was affected by exposure to the immunogen. Whereas immunochallenge did not influence whether foraging mice gained information on patch quality while foraging, it led to reductions in predator avoidance during foraging, suggesting that the proportion of space used by foraging mice may be greater as a result of immunochallenge. This increased space use is predicted to increase encounter rates with patchily distributed LD vectors (ticks) and with incidental prey items. 6. Thus, immunochallenge in white-footed mice, and potentially pathogenic infection, have the potential to indirectly impact community interactions, including those important for pathogen transmission.

Rickettsia felis, West Indies

To the Editor: A spay–neuter (sterilization) program for feral cats from Basseterre, the capital of the Caribbean Island St. Kitts, found that most (45/58; 66%) cats had antibodies to spotted fever group rickettsiae (SFGR). The antibodies were detected with Rickettsia rickettsii antigen in a standard microimmunofluorescence assay (1). Titers for 13 (20%) cats were >320. Most SFGR are transmitted by ticks, but because of their grooming habits, cats seldom have many ticks (2), and we did not find any ticks on the cats we saw through the program. We did, however, commonly find cat fleas, Ctenocephalides felis, which are the main vector of R. felis, a recently described member of the SFGR. R. felis seems to be apathogenic in cats (3) but is the agent of flea-borne spotted fever in humans (4). Although R. felis has been reported from North and South America, Europe, Africa, the Middle-East, and Oceania (4), its presence in the Caribbean islands has not been established. To provide this information we tested DNA extracted with the QIAamp DNA Mini-Kit (QIAGEN, Valencia, CA, USA) from C. felis fleas preserved in 70% ethanol. Of 57 (19%) C. felis fleas from St. Kitts, 11 were positive for R. felis DNA when tested by PCR using primers targeting SFGR ompA (5) or TaqMan assay using primers targeting gltA and a probe specific for the organism (6,7). For a negative control we used distilled water; for a positive control we used DNA from R. montanensis cultures or recombinant control plasmids constructed by amplifying target fragments from R. typhi strain Wilmington and R. felis strain LSU (7). The sequences of the ompA and gltA amplicons obtained had 100% nucleotide sequence similarity with homologous fragments of the type reference isolate R. felis URRxCal2. We used the National Center for Biotechnology Information basic local alignment sequence tool, BLAST (www.ncbi.nlm.nih.gov/blast/Blast.cgi). To determine whether R. felis occurs on another Caribbean island, we tested 32 C. felis fleas from Dominica and found 1 (3%) to be positive by PCR when primers targeting ompA were used. The sequence obtained was also identical to that of R. felis URRxCal2. Our study provides further evidence that cats can be sentinels for the presence of rickettsiae (1). However, although rickettsemia can develop in cats experimentally infected with R. felis (3), no compelling evidence shows that cats help maintain the organism or transmit it to humans (8,9). Rather, it appears that C. felis fleas, which are also commonly found on dogs and to a lesser extent other mammals, are the major reservoir hosts and vectors of infection, although the exact mechanisms are unknown (10). Our study also expands the known distribution of R. felis and should alert healthcare workers who see residents of or vacationers from the Caribbean islands of the possibility of flea-borne spotted fever in their patients.

Ixodes ricinus ticks are reservoir hosts for Rickettsia helvetica and potentially carry flea-borne Rickettsia species

BackgroundHard ticks have been identified as important vectors of rickettsiae causing the spotted fever syndrome. Tick-borne rickettsiae are considered to be emerging, but only limited data are available about their presence in Western Europe, their natural life cycle and their reservoir hosts. Ixodes ricinus, the most prevalent tick species, were collected and tested from different vegetation types and from potential reservoir hosts. In one biotope area, the annual and seasonal variability of rickettsiae infections of the different tick stages were determined for 9 years.ResultsThe DNA of the human pathogen R. conorii as well as R. helvetica, R. sp. IRS and R. bellii-like were found. Unexpectedly, the DNA of the highly pathogenic R. typhi and R. prowazekii and 4 other uncharacterized Rickettsia spp. related to the typhus group were also detected in I. ricinus. The presence of R. helvetica in fleas isolated from small rodents supported our hypothesis that cross-infection can occur under natural conditions, since R. typhi/prowazekii and R. helvetica as well as their vectors share rodents as reservoir hosts. In one biotope, the infection rate with R. helvetica was ~66% for 9 years, and was comparable between larvae, nymphs, and adults. Larvae caught by flagging generally have not yet taken a blood meal from a vertebrate host. The simplest explanation for the comparable prevalence of R. helvetica between the defined tick stages is, that R. helvetica is vertically transmitted through the next generation with high efficiency. The DNA of R. helvetica was also present in whole blood from mice, deer and wild boar.ConclusionBesides R. helvetica, unexpected rickettsiae are found in I. ricinus ticks. We propose that I. ricinus is a major reservoir host for R. helvetica, and that vertebrate hosts play important roles in the further geographical dispersion of rickettsiae.

A cluster-randomised intervention trial against Schistosoma japonicum in the Peoples' Republic of China: bovine and human transmission.

BackgroundZoonotic schistosomiasis japonica is a major public health problem in China. Bovines, particularly water buffaloes, are thought to play a major role in the transmission of schistosomiasis to humans in China. Preliminary results (1998–2003) of a praziquantel (PZQ)-based pilot intervention study we undertook provided proof of principle that water buffaloes are major reservoir hosts for S. japonicum in the Poyang Lake region, Jiangxi Province.Methods and FindingsHere we present the results of a cluster-randomised intervention trial (2004–2007) undertaken in Hunan and Jiangxi Provinces, with increased power and more general applicability to the lake and marshlands regions of southern China. The trial involved four matched pairs of villages with one village within each pair randomly selected as a control (human PZQ treatment only), leaving the other as the intervention (human and bovine PZQ treatment). A sentinel cohort of people to be monitored for new infections for the duration of the study was selected from each village. Results showed that combined human and bovine chemotherapy with PZQ had a greater effect on human incidence than human PZQ treatment alone.ConclusionsThe results from this study, supported by previous experimental evidence, confirms that bovines are the major reservoir host of human schistosomiasis in the lake and marshland regions of southern China, and reinforce the rationale for the development and deployment of a transmission blocking anti-S. japonicum vaccine targeting bovines.Trial RegistrationAustralian New Zealand Clinical Trials Registry ACTRN12609000263291

Gammaherpesvirus carrier status of black wildebeest (Connochaetes gnou) in South Africa

Malignant catarrhal fever (MCF) is an economically important disease primarily of domestic cattle with a high case fatality rate. It is caused by either alcelaphine herpesvirus type 1 (AlHV-1) or ovine herpesvirus type 2 (OvHV-2). The major reservoir host of AlHV-1 is the blue wildebeest (Connochaetes taurinus), but it is generally accepted that the black wildebeest (Connochaetes gnou) is also a reservoir host. No viral studies in the black wildebeest have been reported and the carrier status of black wildebeest has not been documented. Specimens were collected from several game farms and conservation areas in central South Africa representing the geographical area historically linked to the natural habitat of the black wildebeest. Specimens were obtained from 304 black wildebeest of different ages and sex, as well as 51 black wildebeest foetuses at different stages of gestation. Virus was isolated from a black wildebeest calf. Morphological features and antigenic characteristics suggested it to be a gammaherpesvirus closely related to AlHV-1. All serum samples tested positive with a competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA) for group-specific malignant catarrhal fever virus antibody. A SYBR Green real-time PCR assay was developed for the detection of gammaherpesviral DNA. Only 15.8 % of the animals tested positive with the real-time PCR assay whereas 90 % of the foetuses tested positive. This finding suggests that, unlike OvHV-2 infection in lambs in which the infection takes place after weaning, the virus in black wildebeest is mainly transmitted in utero or soon after birth. The results suggest that black wildebeest are latent carriers of a gammaherpesvirus similar or closely related to AlHV-1 present in blue wildebeest and that it is likely that all black wildebeest are persistently infected.

Isolation and Molecular Detection of Ehrlichia from Vertebrate Animals

Human monocytic ehrlichiosis (HME), caused by Ehrlichia chaffeensis, was first recognized in 1986. Infection with this pathogen can be fatal in immune compromised and elderly humans. E. chaffeensis can also infect dogs and several wild animals. The clinical symptoms of HME include fever, headache, malaise, myalgia, confusion, rash, lymphadenopathy, and nausea. White-tailed deer serve as the major reservoir host for the natural maintenance of E. chaffeensis. E. canis is primarily responsible for the canine monocytic ehrlichiosis and is endemic throughout the world. It has a significant impact on the health of dogs. The isolation and growth of Ehrlichia species from vertebrate host samples is difficult and time consuming. In this unit, methods to recover E. chaffeensis and E. canis from infected blood samples collected from dogs, deer, and human patients are described. PCR and RT-PCR methods for sensitive detection of Ehrlichia infection are also discussed.

A Novel Exo-antigen-based ELISA for the Detection of Canine Leishmaniasis

Dogs which are infected with leishmania parasites serve as major reservoir hosts for zoonotic visceral leishmaniasis. The incidence of zoonotic visceral leishmaniasis is rising in many countries. This may be associated with the continuing drift of people and their pets from rural areas into peri-urban settings, particularly at the fringe of large cities. At the same time, there is evidence of adaptation of sand fly vectors to these urban settings. This has created an alarming situation because, even though domestic and stray dogs may be infected, many remain asymptomatic but are still highly infectious to the sand fly vectors and thus pose a serious threat to human health. Over half of the infected dogs have asymptomatic infections and current assays are not sensitive enough under field conditions to distinguish asymptomatic from symptomatic dogs. There is an urgent need for a specific and sensitive screening tool for use in the field. We have previously demonstrated that promastigote exo-antigen-based ELISAs can be used in the specific diagnosis of human visceral leishmaniasis (HVL). A cocktail of exo-antigens prepared from three species (L. infantum, L. donovani, and L. major) was used to develop and optimize a canine ELISA assay. Serum samples from dogs with a variety of pathological conditions but living in a non-leishmania endemic area were used as negative controls and their reactivity was used to determine a cut-off value for the ELISA. Samples from dogs residing in a leishmania endemic area were tested in parallel using direct agglutination (DAT), immunofluorescence (IFAT), and ELISA. The ELISA results correlated closely (100%) with the clinical symptoms, and were elevated in one asymptomatic dog. This sample was also found to be positive by IFAT. Based on its sensitivity and specificity, the cocktail exo-antigen-based ELISA may prove useful, even at 1:2,000 serum dilutions, for screening dogs in different geographical regions of the world.