A new stable focus of canine leishmaniasis in northern Italy

A new stable focus of canine leishmaniasis (CanL) was identified in a coastal Adriatic area of the Emilia-Romagna Region in Northern Italy. Following the first clinical cases observed starting from 1998, a seroepidemiological survey was carried out on owned dogs from two communes and on animals housed in dog pounds of the Rimini province. Sixteen out of 612 dogs (2.6%) resulted positive to the IFA test. The 16 positive dogs all came from the two communes, with seroprevalences of 3 and 6%, respectively. The autochthonous origin of the infection was confirmed in all the cases. The parasitological investigation led to the isolation and identification of the parasite as Leishmania infantum Zymodeme MON 1. An entomological survey showed that Phlebotomus perniciosus and P. perfiliewi are present in this area and that P. perfiliewi was very abundant in one collection site. The risk of the establishment of a permanent transmission of the infection in the area, previously considered CanL-free, must be analysed in view of further investigations to be extended also to neighbouring areas.     

Canine leishmaniasis in the Rif mountains (Moroccan Mediterranean coast): a seroepidemiological survey

A sero-epidemiological survey has been conducted in several localities of the province of Nador to investigate canine leishmaniasis in the North-Eastern slope of the Rif mountains (Mediterranean coast of Morocco). Serum samples collected from 257 dogs were analysed using indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) to detect anti-Leishmania infantum antibodies. Forty eight (18.7%) of the screened dogs were IFAT positive and 54 (21.0%) were ELISA positive; the concordance of the two methods was 96.1%. The prevalence of infection is significantly higher in dogs more than four years of age whereas no significant difference in prevalence of infection was seen between males and females. The frequent symptoms observed in seropositive dogs were the enlargement of lymph nodes (57.4%), emaciation (51.9%) and skin involvement (25.9%). However, 38.9% of those dogs showed no one of the major symptoms of visceral leishmaniasis. Leishmania isolated from three of the examined dogs was identified as L. infantum MON-1. These results show that the North-Eastern slope of the Rif mountains is one of the most active Mediterranean areas of visceral leishmaniasis and confirm that the dog is the main reservoir of L. infantum.     

Visceral leishmaniasis in eastern Sudan: parasite identification in humans and dogs; host-parasite relationships

In 1996, an epidemic outbreak of visceral leishmaniasis (VL) started in Barbar el Fugara, a village in Gedarif State (eastern Sudan). From 1997 to 2000, regular epidemiological studies were carried out in the human population, as well as in mammals and sand flies. In symptomatic patients, 46/69 lymph node, 6/20 post kala-azar dermal leishmaniasis (PKDL) and 1/4 cutaneous cultures in NNN medium were positive. In 69 dogs, 23/79 lymph node cultures were positive. In other mammals (47 rodents, five donkeys, one mongoose and one monkey) spleen and/or blood cultures were negative. Characterization of isolated strains (by starch gel electrophoresis and isoelectrofocusing) identified three zymodemes of Leishmania donovani, two of L. infantum and two of L. archibaldi complexes from patient samples and three zymodemes of L. donovani, three of L. infantum and two of L. archibaldi complexes from dog samples. Five of them were present in both man and dog. For the first time, a strain from a PKDL case was identified as L. infantum, and a child had the same L. infantum zymodeme in VL and in subsequent PKDL. Blood samples from dogs were studied by immunofluorescent antibody test (IFAT). The seroprevalence in dogs was 72.5%, 74.3% and 42.9% in 1998, 1999 and 2000, respectively. By using CDC miniature light traps 12 745 sand flies were collected and then identified. Phlebotomus papatasi (7%) and P. orientalis (5%) were sympatric, mainly inside homes (85% and 75%, respectively). These results, the relative stability of seroprevalence in dogs and the intradomiciliar presence of P. orientalis, known as a vector of VL in Sudan, suggest several hypotheses: (i) man is responsible for the disease in dogs, (ii) the dog is the reservoir of VL, (iii) the dog is an intermediate host between a possible sylvatic cycle and the anthroponotic cycle. More extensive studies are needed to assess the transmission cycle of VL in this area of Sudan.     

A stable focus of canine leishmaniosis in the Bologna Province, Italy

During an epidemiological survey carried out for two consecutive years (2001-2002), autochthonous cases of canine leishmaniosis (CanL) were reported in Communes of the Bologna Province (Emilia-Romagna Region, northern Italy), involved in the past (1971-1972) in a severe outbreak of human visceral leishmaniosis (VL). Serological controls, carried out by immunofluorescence antibody test on a sample of owned dogs, detected a mean prevalence of 2.5% in the first year in 4 Communes, and of 11.2% in the second year in only one Commune, where an incidence value of 9.3% was assessed. The autochthonous origin of the infection was confirmed in 11 out of 13 positive animals in the first year and in 5 out of 6 new cases in the second year. In one case the parasitological examinations led to the isolation of leishmaniae characterized as Leishmania infantum zymodeme MON 1. Entomological surveys carried out during two sandfly seasons in one of the areas concerned by the VL outbreak showed the presence of Phlebotomus perfiliewi and, to a lesser extent, of P perniciosus, both proven vectors of L. infantum in Italy. The results obtained seem to suggest the presence of a stable focus of CanL in the territory involved in the previous VL outbreak of 1971-1972, within which the infection in the canine population had been assessed only serologically. Such an epidemiological situation may be seen either as the persistence of an old focus or as a new imported one.     

Detection of Leishmania infantum in naturally infected Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae) and Canis familiaris in Misiones, Argentina: the first report of a PCR-RFLP and sequencing-based confirmation assay

In this study, a genotypification of Leishmania was performed using polimerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing techniques to identify species of Leishmania parasites in phlebotomine sand flies and dogs naturally infected. Between January-February of 2009, CDC light traps were used to collect insect samples from 13 capture sites in the municipality of Posadas, which is located in the province of Misiones of Argentina. Sand flies identified as Lutzomyia longipalpis were grouped into 28 separate pools for molecular biological analysis. Canine samples were taken from lymph node aspirates of two symptomatic stray animals that had been positively diagnosed with canine visceral leishmaniasis. One vector pool of 10 sand flies (1 out of the 28 pools tested) and both of the canine samples tested positively for Leishmania infantum by PCR and RFLP analysis. PCR products were confirmed by sequencing and showed a maximum identity with L. infantum. Given that infection was detected in one out of the 28 pools and that at least one infected insect was infected, it was possible to infer an infection rate at least of 0.47% for Lu. longipalpis among the analyzed samples. These results contribute to incriminate Lu. longipalpis as the vector of L. infantum in the municipality of Posadas, where cases of the disease in humans and dogs have been reported since 2005.     

Time-activity budget of urban-adapted free-ranging dogs

The domestic dog (Canis lupus familiaris) is known to have evolved from gray wolves, about 15,000 years ago. They frequently exist as free-ranging populations across the world. They are typically scavengers and well adapted to living among humans. Most canids living in and around urban habitats tend to avoid humans and show crepuscular activity peaks. In this study, we carried out a detailed population-level survey on free-ranging dogs in West Bengal, India, to understand the activity patterns of free-ranging dogs in relation to human activity. Using 5669 sightings of dogs, over a period of 1 year, covering the 24 h of the day, we carried out an analysis of the time activity budget of free-ranging dogs to conclude that they are generalists in their habit. They remain active when humans are active. Their activity levels are affected significantly by age class and time of the day. In addition, we provide a detailed ethogram of free-ranging dogs. This, to our knowledge, is the first study of this kind, which might be used to further study the eco-ethology of these dogs.

     

Secreted antigens of the amastigote and promastigote forms of Leishmania infantum inducing a humoral response in humans and dogs.

To study the antigens secreted by promastigote and amastigote forms of Leishmania infantum which are able to induce a humoral response in human patients and dogs, we have carried out immunoprecipitation assays with different supernatants of in vitro cultured parasites, metabolically labelled with [35S]methionine, using serum samples from human patients and dogs. In addition, some metabolic labelling experiments were performed daily during the in vitro culture parasite's life cycle to follow the time course excretion-secretion of parasitic antigens. The results demonstrated that the two different hosts developed an antibody response against secreted antigens of both stages of Leishmania infantum. Nevertheless, the humoral response directed against the excreted-secreted antigens of the promastigote forms was qualitatively and quantitatively different when we compare the human and the dog immune responses. On the other hand, when the excreted-secreted antigens of the amastigote forms are immunoprecipitated with either human or canine immune serum, the humoral response is similar. In addition, the time course study showed that excretion-secretion of antigens was qualitatively and quantitatively modulated during the parasitic in vitro life cycle.     

Analysis of renal immune-deposits in canine leishmaniasis. Preliminary results

Previous studies carried out on 34 dogs spontaneously infected by Leishmania infantum showed the presence of kidney lesions characterized by immunologically mediated glomerular and tubular damage. Glomerular immune-deposits were studied in 13 of these dogs. Immunoglobulins were isolated from kidney tissues by acid elution; IgG fractions from eluates, obtained by ammonium sulfate precipitation, were subjected to clonotypic analysis by autoradiography after isoelectrofocusing (IEF) using 125I radiolabelled goat IgG fraction-anti Fab2 of dog IgG. Idiotypic characterization of IgG eluted from kidney tissues was performed by IEF and autoradiography using both 125I radiolabelled membrane antigens of L. infantum extracted by Triton x 100 and 125I radiolabelled dog IgG for rheumatoid or anti-idiotypic activity. The IgG deposited in the kidney tissues of examined dogs were polyclonal and a specific activity against Leishmania membrane antigens was revealed. Meanwhile an anti-IgG activity of deposited immunoglobulins was not observed.     

A serological and epidemiological evaluation of risk factors for toxocariasis in children in central Poland

The level of specific anti-Toxocara IgG was examined in 343 children from central Poland with suspected Toxocara infection. Based on the presence of specific IgG antibodies and clinical signs and symptoms, toxocariasis was confirmed in 249 patients. The serological results of diagnosed cases were analysed in connection with epidemiological data (contamination of soil around the children's homes and presence of a dog or a cat). A higher prevalence of soil contamination with Toxocara eggs was found in suburban and rural areas (30.9% and 24.6%, respectively) whereas it was lower in urban areas (10.3%). In about 40% of the children whose households were not contaminated, a decrease in the antibody level was observed 2 years after treatment for toxocariasis, while there was no such decline in those living in contaminated places. The logistic regression model employed to determine the correlation between seropositivity in the children and selected epidemiological risk factors showed a statistically significant relationship in connection only with dog ownership (P = 0.0238). The present results have demonstrated the high risk of toxocariasis for children from rural and suburban areas in Poland. The sero-epidemiological investigations indicated that re-infection might be the reason for persistence of seropositivity after treatment.     

Occupational risk of cutaneous larva migrans: A case report and a systematic literature review

Cutaneous larva migrans (CLM) is a parasitic zoonosis of warm tropical and subtropical areas, although autochthonous cases have been increasingly reported in Western European countries. Data on the prevalence of CLM as an occupational disease in workers exposed to potentially contaminated soil or in close contact with dogs and cats are scant. Herein, we report an autochthonous case of CLM in a dog breeder from southern Italy (Apulia region), along with a systematic literature review describing the risk of CLM infection, mainly according to job categories. The patient was referred to the dermatology unit presenting a serpiginous lesion on his hand, raising the suspected CLM diagnosis. In non-endemic areas, CLM might represent a challenge for physicians in terms of diagnosis, treatment, and prevention, particularly in workplaces. The multidisciplinary approach in the diagnosis of CLM with the involvement of different scientific competences (i.e., dermatologists, veterinarians, and occupational physicians) may contribute to further assess the distribution of human CLM and associated risk factors, toward reducing the risk for the infection.     

The companion dog as a model for human aging and mortality

Around the world, human populations have experienced large increases in average lifespan over the last 150 years, and while individuals are living longer, they are spending more years of life with multiple chronic morbidities. Researchers have used numerous laboratory animal models to understand the biological and environmental factors that influence aging, morbidity, and longevity. However, the most commonly studied animal species, laboratory mice and rats, do not experience environmental conditions similar to those to which humans are exposed, nor do we often diagnose them with many of the naturally occurring pathologies seen in humans. Recently, the companion dog has been proposed as a powerful model to better understand the genetic and environmental determinants of morbidity and mortality in humans. However, it is not known to what extent the age‐related dynamics of morbidity, comorbidity, and mortality are shared between humans and dogs. Here, we present the first large‐scale comparison of human and canine patterns of age‐specific morbidity and mortality. We find that many chronic conditions that commonly occur in human populations (obesity, arthritis, hypothyroidism, and diabetes), and which are associated with comorbidities, are also associated with similarly high levels of comorbidity in companion dogs. We also find significant similarities in the effect of age on disease risk in humans and dogs, with neoplastic, congenital, and metabolic causes of death showing similar age trajectories between the two species. Overall, our study suggests that the companion dog may be an ideal translational model to study the many complex facets of human morbidity and mortality.     

Canine filariosis in Umbria: an update of the occurrence one year after the first observation of autochthonous foci

Following the first observation of two autochthonous foci of canine filariosis occurred in Umbria region in the year 2001, a survey on prevalence and risk factors was conducted 12 months later to better understand the actual entity of the Dirofilaria problem in Umbria region. Blood samples were collected between January and December 2002 from 2406 dogs living in a total of 7 towns located in the identified areas at risk. Blood samples were tested by a modified Knott's technique to evaluate the microfilaraemia and, by a commercial ELISA kit, to detect in the sera adult antigens of D. immitis. The results were subject to statistical analysis. A total of 439 dogs were found to be infected. The true prevalence (LC 95%) was of 18%. Microfilariae of D. immitis were detected in 286 dogs (13%) while 112 dogs (6%) showed only microfilariae of D. repens and 41 dogs (1.6%) microfilariae of both D. immitis and D. repens. The prevalence ratio (PR) for each species of Dirofilaria (LC 95%) calculated in association with different risk factors (age, sex, use, outdoor night status, position, living together with other dogs, breed) and the statistical significance between the risk factors and the presence/absence of the infection, evaluated for each species of Dirofilaria, are discussed.     

Susceptibility to visceral leishmaniasis in the domestic dog is associated with MHC class II polymorphism

Zoonotic visceral leishmaniasis (VL) is a disease of dogs, humans and other animals caused by the intracellular macrophage parasite Leishmania infantum. We examined the relationship between DLA class II alleles ( DRB1, DQA1, DQB1) and the course of infection in a cohort of Brazilian mongrel dogs exposed to natural L. infantum infection. DLA alleles were typed by sequence-based typing. DLA-DRB1 genotype was significantly associated with levels of anti- Leishmania IgG and parasite status assessed by PCR. Dogs with DLA-DRB1*01502 had higher levels of specific IgG and an increased risk of being parasite positive compared with dogs without this allele, controlling for other alleles and significant variables. No significant associations were seen for DLA-DQA1 or DLA-DQB1 alleles. These results suggest that the DLA-DRB1 locus plays a role in determining susceptibility to canine VL. As the domestic dog is the main reservoir for human infection, the identification of genetic factors influencing canine resistance or susceptibility to VL may provide insights into the immunology and potential control through vaccination of VL.     

Monitoring of canine leishmaniasis in northern Italy: an update from a scientific network

Canine leishmaniasis (CanL) due to Leishmania infantum is a disease of great veterinary importance and a serious public health problem. In humans, L. infantum causes visceral (VL) and cutaneous leishmaniasis (CL) and the distribution of VL overlaps that of CanL. Currently, VL is considered by WHO as an emerging zoonosis in southern Europe. The dog is the only domestic reservoir of the infection and phlebotomine sandflies are the only proven vectors of leishmaniasis for dogs and humans. CanL is endemic in Italy, particularly in central and southern regions, including islands. Until 1983, all regions of northern Italy but Liguria and some territories of Emilia Romagna were considered free from CanL. From early '90s new stable foci of CanL have appeared, most of them located within classical endemic areas including territories of Emilia Romagna, Tuscany, Umbria, Marche, and Abruzzi regions. But the most relevant aspect, from an epidemiological point of view, has been the appearance of stable CanL foci in northern Italy, namely in Veneto and Piedmont regions. In these two foci, entomological surveys showed the presence of P. perniciosus and of a second phlebotomine vector, P. neglectus, which may have played a role in the CanL diffusion in some parts of northern Italy. Furthermore, in these areas, autochthonous human VL cases have occurred. There is therefore a realistic risk that CanL infection could rapidly spread through northern latitudes and a surveillance activity is strongly needed. For this reason, in October 2002, thanks to the collaboration and support of Intervet Italia, the network "LeishMap" was created, with the main purpose of monitoring the spread of CanL and vectors in northern Italy. LeishMap consists of scientific and sanitary institutions with proven experience both in field surveys and diagnostic methodologies on CanL and phlebotomine vector. It is organised in 4 Operational Units (OU), represented by researchers of the Veterinary Faculties of the University of Bologna, Padua, Milan and Turin, under the scientific coordination of the MIPI Department, ISS of Rome and with the collaboration of private and public veterinarians operating in the regions under study. During the first year of activity, each OU was involved in the serological and entomological surveillance of several territories in the respective regions, where recent autochthonous CanL cases were registered. The studies have involved five regions, namely Valle D'Aosta, Piedmont, Lombardia, Veneto, Trentino-Alto Adige and Emilia Romagna. In the Symposium 6 of this Congress we report detailed results of a retrospective analysis of data concerning CanL and vectors in northern Italy till 2002 and the preliminary results of 2003 on the seroprevalence rates observed in foci studied and on the entomological surveys carried out. In summary, the results outlined that already known foci of CanL are expanding from the original sites. Several new foci have been identified and many others are at high risk of evolving toward a stable endemicity. P. perniciosus has been found in all but one the suspected new foci. In Emilia Romagna region P. perfiliewi was identified in 2 areas and in one was the only species present. The occurrence of P. neglectus was confirmed in three regions, Veneto, Lombardia and Piedmont. In conclusion, from the 2002-2003 LeishMap activities it appears that further monitoring activities are necessary to identify new endemic foci of CanL, this representing the prerequisite for the implementation of programs for leishmaniasis control in northern Italy.     

Transplacental transmission of a North American isolate of Leishmania infantum in an experimentally infected beagle

Leishmania infantum, an etiologic agent of zoonotic visceral leishmaniasis, is widespread among foxhounds in the United States. Although sand flies are widely distributed throughout the United States, epidemiological data do not support a major role for sand flies in the transmission of L. infantum in foxhounds in this country. Congenital transmission of human visceral leishmaniasis is reported in humans and might also occur in dogs. We have previously isolated L. infantum from Virginia foxhounds and used this isolate (LIVT-1) to experimentally infect beagles. Four female beagles, chronically infected with LIVT-1, were bred to a male beagle chronically infected with L. infantum chagasi. One beagle was able to maintain her pregnancy, and 4 puppies were delivered by cesarean section. One puppy was malformed and autolytic at delivery, and tissues were not collected or analyzed. The remaining puppies were killed at the time of cesarean section, and selected tissues were collected for parasite culture and PCR. Promastigotes were not cultured from tissues in any of the puppies. Leishmania sp. DNA was detectable by PCR in liver, bone marrow, and heart from all 3 puppies and in the spleen, lymph node, kidney, and placenta in 2 puppies. Placental tissue from the dam was PCR negative. This is the first report of maternal transmission of a North American isolate of L. infantum from an experimentally infected dog.     

Feline leishmaniasis: what's the epidemiological role of the cat?

Feline leishmaniasis (FL) is a quite uncommon feature. Clinical disease has been described in cats since nineties begin. More than 40 reports in world literature have been referred, but the clinical cases have been only recently well defined. Most of the reports focus on infected cats living in endemic areas, even if, more recently FL due to Leishmania infantum was found in Sao Paulo State, in Brazil where autochthonous human or canine leishmaniasis cases have never reported. In Europe clinical cases of FL have been described from Portugal, France, Spain and Italy from 1996 to 2002. When a typing of the etiological agent was performed L. infantum was identified in all reported cases. In some endemic areas serological surveys have also been carried out in cats, using IHAT in Egypt, Western blot in France or IFAT in Italy. Sixty Egyptian cats had low serological antibody titers, from 1/32 to 1/128, in the endemic focus of canine leishmaniasis of Alpes Maritimes 12 out of 97 (12.5%) cats showed antibodies versus antigens 14 and/or 18 kDa of L. infantum. A previous survey by means of IFAT in Liguria and Toscana on 110 and 158 feline sera respectively reports a seroprevalence of 0.9% with low titer, while sera from Sicily seem to be positive at higher dilutions. Animals living in an endemic area can develop specific antibodies against leishmania and, in our experience, they can be evidentiated by means of IFAT. The antibody titers appear to be lower in affected cats than in dogs, even if the number of clinical cases is very scanty. PCR tests on feline blood samples are in progress, but preliminary results confirm the presence of leishmania DNA in such specimens. Cutaneous leishmaniasis is the more frequent form in cats and it was reported from several countries. Typical signs include nodular to ulcer or crusty lesions on the nose, lips, ears, eyelids, alopecia: clinical signs of cutaneous FL are unspecific and in endemic area this infection must be taken into account. Visceral leishmaniasis is not common in cats: this form shows visceral involvement: liver and spleen are interested, with lymph nodes and kidney. The cat probably has to considerate to play an active role in the disease, in contrast to goats, calves and horses who could act as accidental reservoirs of leishmania, while sheep appears to be not susceptible to experimental infection. In endemic foci for kala-azar in Sudan cows, goats and donkeys had a high prevalence of specific antibodies. Recently in Europe sporadic cases of equine leishmaniasis have been reported: L. infantum was the causative agent. Equine leishmaniasis appears as a self-healing skin-dwelling disease, with a massive accumulation of parasites. The animals do not often show detectable specific antibodies and recover without any chemotherapy. Untreated affected cats can frequently die and we also observed lymph nodes and blood involvement indicating a spread of leishmania in feline hosts. The epidemiological role of the cat has never been clarified due also to lack of xenodiagnosis trials. This species is believed to have a high degree of natural resistance, as observed following experimental infection. Some of the affected cats were FIV and/or FeLV positive and these viroses such as stress may induce an impaired cellular immune response, even if leishmania infected cat was not submitted to CD4+, CD8+ lymphocyte counts nor other immunological test. However the resistance of the cat to leishmania infection probably depends on genetic factors, not strictly related to cell mediated immunity, taking into account the high seroprevalence of FIV infections (30%) in our country versus the number of clinical cases.     

Factors Associated with the Seroprevalence of Leishmaniasis in Dogs Living around Atlantic Forest Fragments

Canine visceral leishmaniasis is an important zoonosis in Brazil. However, infection patterns are unknown in some scenarios such as rural settlements around Atlantic Forest fragments. Additionally, controversy remains over risk factors, and most identified patterns of infection in dogs have been found in urban areas. We conducted a cross-sectional epidemiological survey to assess the prevalence of leishmaniasis in dogs through three different serological tests, and interviews with owners to assess features of dogs and households around five Atlantic Forest remnants in southeastern Brazil. We used Generalized Linear Mixed Models and Chi-square tests to detect associations between prevalence and variables that might influence Leishmania infection, and a nearest neighbor dispersion analysis to assess clustering in the spatial distribution of seropositive dogs. Our findings showed an average prevalence of 20% (ranging from 10 to 32%) in dogs. Nearly 40% (ranging from 22 to 55%) of households had at least one seropositive dog. Some individual traits of dogs (height, sterilization, long fur, age class) were found to positively influence the prevalence, while some had negative influence (weight, body score, presence of ectoparasites). Environmental and management features (number of cats in the households, dogs with free-ranging behavior) also entered models as negative associations with seropositivity. Strong and consistent negative (protective) influences of the presence of chickens and pigs in dog seropositivity were detected. Spatial clustering of cases was detected in only one of the five study sites. The results showed that different risk factors than those found in urban areas may drive the prevalence of canine leishmaniasis in farm/forest interfaces, and that humans and wildlife risk infection in these areas. Domestic dog population limitation by gonadectomy, legal restriction of dog numbers per household and owner education are of the greatest importance for the control of visceral leishmaniasis in rural zones near forest fragments.     

Borrelia burgdorferi in ticks and dogs in the province of Vojvodina, Serbia

Lyme disease is a tick borne zoonotic infection, caused by Borrelia burgdorferi s.l. bacteria. For the transmission of the disease, the presence of ticks is a prerequisite. Lyme borreliosis mostly occurs in people and dogs, but it may occur in other animals. Ticks which carry B. burgdorferi s.l. in Serbia are of the Ixodes ricinus specis. In Serbia, Lyme disease was detected for the first time in the late '80-es. In dogs, clinical symptoms may occur even months after a tick bite, and include weakness, lymphadenopathy, fever, lameness, arthritis, etc. In our survey, we have observed tick and dog populations in the province of Vojvodina (northern part of Serbia). I. ricinus ticks were collected and examined for the presence of B. burgdorferi s.l. in several chosen locations. In addition, blood samples were collected from house dogs and pets from the same locations, and analyzed for the presence of antibodies specific for B. burgdorferi s.l. The results showed a mean infection of ticks of 22.12%, and a mean seroprevalence of Lyme disease in dogs of 25.81%. We conclude that in Vojvodina there is an actual risk of Lyme borreliosis for other animals and humans, because of the persistence of B. burgdorferi s.l. in both tick and dog populations.     

Role of dog behaviour and environmental fecal contamination in transmission of Echinococcus multilocularis in Tibetan communities

On the Eastern Tibetan Plateau region (Sichuan province, China) dogs are regarded as important definitive hosts of Echinococcus multilocularis. We studied dog spatial behaviour in 4 Tibetan villages in order to determine the role of dogs in environmental contamination and their potential interactions with small mammal intermediate hosts. We identified definitive host species and Echinococcus spp. infection status of feces collected in the field by PCR methods and analysed the spatial distribution of canid feces. Nocturnal space utilization of GPS collared dogs in and around villages was also undertaken. E. multilocularis DNA was amplified in 23% of dog feces (n=142) and in 15% of fox feces (n=13) but this difference was not significant. However, dog feces were more frequently observed (78% of collected feces) than fox feces and are therefore assumed to largely contribute to human environment contamination. Feces were mainly distributed around houses of dog owners (0-200 m) where collared dogs spent the majority of their time. Inside villages, the contamination was aggregated in some micro-foci where groups of dogs defecated preferentially. Finally, small mammal densities increased from the dog core areas to grasslands at the periphery of villages occasionally used by dogs; male dogs moving significantly farther than females. This study constitutes a first attempt to quantify in a spatially explicit way the role of dogs in E. multilocularis peri-domestic cycles and to identify behavioural parameters required to model E. multilocularis transmission in this region.     

Toxigenic Corynebacterium ulcerans isolated from a hunting dog and its diphtheria toxin antibody titer

Toxigenic Corynebacterium ulcerans is a zoonotic pathogen that produces diphtheria toxin and causes a diphtheria‐like illness in humans. The organism is known to infect and circulate among dogs, which can then transmit it to humans. Furthermore, previous studies have found that C. ulcerans is carried by wild animals, including game animals. In the present study, we tested hunting and companion dogs for the presence of toxigenic C. ulcerans and succeeded in isolating the bacterium from a hunting dog. Moreover, several hunting dogs had serum diphtheria antitoxin titers that were higher than the titers required for protection in humans, suggesting a history of exposure to toxigenic Corynebacterium strains. Notably, ribotyping, pulsed‐field gel electrophoresis and tox gene sequencing demonstrated that the isolate from the hunting dog clustered with previously characterized C. ulcerans strains isolated from wild animals, as opposed to groups of isolates from humans and companion dogs. Interestingly, the wild animal cluster also contains an isolate from an outdoor breeding dog, which could have formed a bridge between isolates from wild animals and those from companion dogs. The results presented herein provide insight into the mechanism by which the zoonotic pathogen C. ulcerans circulates among wild animals, hunting and companion dogs, and humans.