Isolation of Bartonella henselae, Bartonella koehlerae subsp. koehlerae, Bartonella koehlerae subsp. bothieri and a new subspecies of B. koehlerae from free-ranging lions (Panthera leo) from South Africa, cheetahs (Acinonyx jubatus) from Namibia and captive cheetahs from California.

Bartonellae are blood- and vector-borne Gram-negative bacteria, recognized as emerging pathogens. Whole-blood samples were collected from 58 free-ranging lions (Panthera leo) in South Africa and 17 cheetahs (Acinonyx jubatus) from Namibia. Blood samples were also collected from 11 cheetahs (more than once for some of them) at the San Diego Wildlife Safari Park. Bacteria were isolated from the blood of three (5%) lions, one (6%) Namibian cheetah and eight (73%) cheetahs from California. The lion Bartonella isolates were identified as B. henselae (two isolates) and B. koehlerae subsp. koehlerae. The Namibian cheetah strain was close but distinct from isolates from North American wild felids and clustered between B. henselae and B. koehlerae. It should be considered as a new subspecies of B. koehlerae. All the Californian semi-captive cheetah isolates were different from B. henselae or B. koehlerae subsp. koehlerae and from the Namibian cheetah isolate. They were also distinct from the strains isolated from Californian mountain lions (Felis concolor) and clustered with strains of B. koehlerae subsp. bothieri isolated from free-ranging bobcats (Lynx rufus) in California. Therefore, it is likely that these captive cheetahs became infected by an indigenous strain for which bobcats are the natural reservoir.

Attitudes and Beliefs of Pig Farmers and Wild Boar Hunters Towards Reporting of African Swine Fever in Bulgaria, Germany and the Western Part of the Russian Federation.

This study investigated the attitudes and beliefs of pig farmers and hunters in Germany, Bulgaria and the western part of the Russian Federation towards reporting suspected cases of African swine fever (ASF). Data were collected using a web-based questionnaire survey targeting pig farmers and hunters in these three study areas. Separate multivariable logistic regression models identified key variables associated with each of the three binary outcome variables whether or not farmers would immediately report suspected cases of ASF, whether or not hunters would submit samples from hunted wild boar for diagnostic testing and whether or not hunters would report wild boar carcasses. The results showed that farmers who would not immediately report suspected cases of ASF are more likely to believe that their reputation in the local community would be adversely affected if they were to report it, that they can control the outbreak themselves without the involvement of veterinary services and that laboratory confirmation would take too long. The modelling also indicated that hunters who did not usually submit samples of their harvested wild boar for ASF diagnosis, and hunters who did not report wild boar carcasses are more likely to justify their behaviour through a lack of awareness of the possibility of reporting. These findings emphasize the need to develop more effective communication strategies targeted at pig farmers and hunters about the disease, its epidemiology, consequences and control methods, to increase the likelihood of early reporting, especially in the Russian Federation where the virus circulates.

Multivariate analysis of traditional pig management practices and their potential impact on the spread of infectious diseases in Corsica.

Corsica is a French Mediterranean island with traditional extensive pig farming oriented towards the production of high quality cured meat products. The increasing success of these cured products in continental Europe has triggered the development and organisation of an extensive pig farming industry. However, these pig farming practices have seldom been described and analysed to understand the potential risk of introduction and spread of infectious diseases. We conducted a cross-sectional study in Corsica in 2013 to characterise the main pig management practices and to identify groups of farms with similar practices and therefore homogeneous risk of introduction and spread of infectious diseases. We interviewed 68 pig farmers and investigated different farm management practices which could lead to contact between herds, such as trading animals, sharing pastures, feed and reproduction management (direct contacts), slaughtering and carcass waste management, and contacts with people and vehicles (indirect contacts). The practices were described and the farms grouped by multiple factor and hierarchical clustering analyses. Results revealed interesting patterns in the introduction and spread of infectious disease, such as the seasonality of pig production, the potential local spread of diseases in pastures due to the presence of free-ranging boars, carcasses, and animal waste. Multivariate analyses identified four groups of farms with different levels of risk of the spread of infectious disease, illustrating changes in farmers' customs from free-range uncontrolled farming systems to more controlled systems aimed at the production of high quality pork products. These results will be useful to more realistically simulate the spread of infectious diseases among Corsican pig farms and highlight the need for awareness raising campaigns among the stakeholders to reduce risky practices.

How to reach the poor? Surveillance in low-income countries, lessons from experiences in Cambodia and Madagascar.

Surveillance of animal diseases in developing countries faces many constraints. Innovative tools and methods to enhance surveillance in remote and neglected areas should be defined, assessed and applied in close connection with local farmers, national stakeholders and international agencies. The authors performed a narrative synthesis of their own publications about surveillance in Madagascar and Cambodia. They analysed the data in light of their fieldwork experiences in the two countries' very challenging environments. The burden of animal and zoonotic diseases (e.g. avian influenza, African swine fever, Newcastle disease, Rift Valley fever) is huge in both countries which are among the poorest in the world. Being poor countries implies a lack of human and financial means to ensure effective surveillance of emerging and endemic diseases. Several recent projects have shown that new approaches can be proposed and tested in the field. Several advanced participatory approaches are promising and could be part of an innovative method for improving the dialogue among different actors in a surveillance system. Thus, participatory modelling, developed for natural resources management involving local stakeholders, could be applied to health management, including surveillance. Data transmission could benefit from the large mobile-phone coverage in these countries. Ecological studies and advances in the field of livestock surveillance should guide methods for enhancing wildlife monitoring and surveillance. Under the umbrella of the One Health paradigm, and in the framework of a risk-based surveillance concept, a combination of participatory methods and modern technologies could help to overcome the constraints present in low-income countries. These unconventional approaches should be merged in order to optimise surveillance of emerging and endemic diseases in challenging environments.

Avian influenza and Newcastle disease in three risk areas for H5N1 highly pathogenic avian influenza in Mali, 2007-2008.

Our survey aimed to investigate avian influenza (AI) and Newcastle disease (ND) prevalence and risk factors in three areas of Mali at risk for occurrence of H5N1 highly pathogenic avian influenza. Blood samples and cloacal and oropharyngeal swabs were collected from 1470 birds between February 2007 and May 2008 and were tested by commercial enzyme-linked immunosorbent assay to detect antibodies and real-time reverse-transcription (rRT)-PCR to detect virus. Risk factors associated with seropositivity or positive rRT-PCR were identified by random effect logistic regression. AI seroprevalence was significantly lower in birds from commercial farms (0%) than in village backyard birds (3.1%). For backyard birds, no individual risk factors (species, age, sex) were identified, but birds in the Mopti area in the Sahelian zone, where millions of wild birds migrate, were more seropositive than in the Sikasso area in the Sudano-Guinean zone (odds ratio [OR] = 2.0, P = 0.051). Among backyard birds nonvaccinated against ND, ND seroprevalence was 58.4%, and the odds of seropositivity was 2.0 higher in chickens than in ducks, 1.7 higher in females than in males, 3.1 higher in adults than in young birds, and 3.0 higher in poultry from the Sikasso area than from the Mopti area (P < 0.01 in all cases). Prevalence established by rRT-PCR was low for both AI virus (1.1%) and ND virus (2.6%) and was associated with no risk factors for AI but was higher in chickens than in ducks (OR = 5.3, P = 0.05) and in the Sikasso area than in the Mopti area (OR = 3.4, P = 0.027) for ND. For AI and ND, prevalence assessed by serology or rRT-PCR varied over time, although seasonal and interannual variation could not be clearly distinguished. The intracluster correlation coefficient for serologic data was low for AI (0.014) and higher for ND (0.222). These results are useful to optimize surveillance and control strategy for notifiable avian diseases in African countries with similar agroecological and resource-limited contexts.

Emergence of a new genetic lineage of Newcastle disease virus in West and Central Africa--implications for diagnosis and control.

Newcastle disease (ND) is an OIE listed disease caused by virulent avian paramyxovirus type 1 (APMV-1) strains, which affect many species of birds and may cause severe economic losses in the poultry sector. The disease has been officially and unofficially reported in many African countries and still remains the main poultry disease in commercial and rural chickens of Africa. Unfortunately, virological and epidemiological information concerning ND strains circulating in the Western and Central regions of Africa is extremely scarce. In the present study, sequence analysis, pathotyping and detailed genetic characterization of virulent ND strains detected in rural poultry in West and Central Africa revealed the circulation of a new genetic lineage, distinguishable from the lineages described in the Eastern and Southern parts of the continent. Several mismatches were observed in the segment of the matrix gene targeted by the primers and probe designed for the molecular detection of APMV-1, which were responsible for the false negative results in the diagnostic test conducted. Furthermore, deduced amino acid sequences of the two major antigens eliciting a protective immune response (F and HN glycoprotein) revealed protein similarities <90% if compared to some common vaccine strains. Distinct mutations located in the neutralizing epitopes were revealed, indicating the need for detailed assessment of the efficacy of the current vaccines and vaccination practices in Africa. The present investigation provides important information on the epidemiology, diagnosis and control of NDV in Africa and highlights the importance of supporting surveillance in developing countries for transboundary animal diseases.

Risk factors for West Nile virus seropositivity of equids in Guadeloupe.

In Guadeloupe, West Nile virus (WNV) activity was first observed in equids in 2002, and a high seroprevalence was found in 2003. The objective of our study was to determine individual and environmental factors associated with the risk of WNV seropositivity during 2002-2003. Fieldwork was conducted to retrospectively determine the location of equids at the time of virus circulation and to collect information regarding environmental and individual variables. Sera were collected from 369 equids out of an estimated total population of less than 500. Thirty-four environmental and individual variables were investigated. Equids had a higher risk (p<0.001) for WNV seropositivity if they lived within the proximity "distance less than 1.5km" of marshes or swamp forests "a large freshwater formation behind mangroves" or if they remained outside after dusk. Equids living within the proximity of ouassous shrimp (Macrobrachium rosenbergii) basins or sugar cane fields had a lower risk (p<0.001) for WNV seropositivity. These results confirm that WNV circulation is more likely in the humid coastal areas of Guadeloupe. The identification of risk factors is useful for predicting future emergence sites of WNV in the archipelago and other Neotropical islands, and to better target sentinel surveillance in the region.

Prevalence of Bartonella infection in wild African lions (Panthera leo) and cheetahs (Acinonyx jubatus).

Bartonella species are emerging pathogens that have been isolated worldwide from humans and other mammals. Our objective was to estimate the prevalence of Bartonella infection in free-ranging African lions (Panthera leo) and cheetahs (Acinonyx jubatus). Blood and/or serum samples were collected from a convenience sample of 113 lions and 74 cheetahs captured in Africa between 1982 and 2002. Whole blood samples available from 58 of the lions and 17 of the cheetahs were cultured for evidence of Bartonella spp., and whole blood from 54 of the 58 lions and 73 of the 74 cheetahs tested for the presence of Bartonella DNA by TaqMan PCR. Serum samples from the 113 lions and 74 cheetahs were tested for the presence of antibodies against Bartonella henselae using an immunofluorescence assay. Three (5.2%) of the 58 lions and one (5.9%) of the 17 cheetahs were bacteremic. Two lions were infected with B. henselae, based on PCR/RFLP of the citrate synthase gene. The third lion and the cheetah were infected with previously unidentified Bartonella strains. Twenty-three percent of the 73 cheetahs and 3.7% of the 54 lions tested by TaqMan PCR were positive for Bartonella spp. B. henselae antibody prevalence was 17% (19/113) for the lions and 31% (23/74) for the cheetahs. The prevalence of seropositivity, bacteremia, and positive TaqMan PCR was not significantly different between sexes and age categories (juvenile versus adult) for both lions and cheetahs. Domestic cats are thus no longer the only known carriers of Bartonella spp. in Africa. Translocation of B. henselae seronegative and TaqMan PCR negative wild felids might be effective in limiting the spread of Bartonella infection.