Community perception and knowledge of cystic echinococcosis in the High Atlas Mountains, Morocco.

BACKGROUND: Cystic echinococcosis (CE), a neglected zoonosis caused by the larval stage of the tapeworm Echinococcus granulosus, remains a public health issue in many developing countries that practice extensive sheep breeding. Control of CE is difficult and requires a community-based integrated approach. We assessed the communities' knowledge and perception of CE, its animal hosts, and its control in a CE endemic area of the High Atlas Mountains, Morocco. METHODS: We conducted twenty focus group discussions (FGDs) stratified by gender with villagers, butchers and students in ten Berber villages that were purposefully selected for their CE prevalence. RESULTS: This community considers CE to be a severe and relatively common disease in humans and animals but has a poor understanding of the parasite's life cycle. Risk behaviour and disabling factors for disease control are mainly related to cultural practices in sheep breeding and home slaughtering, dog keeping, and offal disposal at home, as well as in slaughterhouses. Participants in our focus group discussions were supportive of control measures as management of canine populations, waste disposal, and monitoring of slaughterhouses. CONCLUSIONS: The uncontrolled stray dog population and dogs having access to offal (both at village dumps and slaughterhouses) suggest that authorities should be more closely involved in CE control. This study also highlights the need for improved knowledge about the transmission cycle of the parasite among communities and health professionals. Inter-sectoral collaboration between health staff, veterinarians, and social scientists appears to be crucial for sustainable control of this parasitic zoonosis.

Documenting the absence of brucellosis in cattle, goats and dogs in a "One Health" interface in the Mnisi community, Limpopo, South Africa.

This study shows the absence of the world's most common bacterial zoonoses caused by Brucella abortus and Brucella melitensis in cattle, goats and dogs in an agro-pastoral community in South Africa, where heifer vaccination against brucellosis with the live Strain 19 vaccine is compulsory. The study site is bordering wildlife reserves with multiple wildlife species infected with brucellosis. The results showed a low seroprevalence (1.4%) in cattle. Seroprevalence in cattle decreased with age after 4 years in females, males were less positive than females and a tissue culture from a brucellin skin test-positive male was negative. The results indicate that Brucella seropositivity in cattle is due to S19 vaccination and not natural infections. This conclusion is reinforced by the absence of Brucella seropositivity in goats (1/593 positive result) and dogs (0/315), which can be seen as potential spillover hosts. Therefore, the close proximity of brucellosis-infected wildlife is not a threat to domestic animals in this controlled setting with vaccination, fencing and movement control.

Spillover of Mycobacterium bovis from wildlife to livestock, South Africa.

During August 2012-February 2013, bovine tuberculosis was detected in communal livestock bordering the Greater Kruger National Park Complex (GKNPC) in South Africa. Using spacer oligonucleotide and variable number tandem repeat typing, we identified the Mycobacterium bovis strain endemic in GKNPC wildlife. Our findings indicate bovine tuberculosis spillover from GKNPC wildlife to neighboring livestock.

Rearing Management and Its Impact on Caseous Lymphadenitis in Sheep.

Two surveys were conducted to assess the prevalence and risk factors of ovine caseous lymphadenitis (CL) and explore the association between its superficial and visceral forms in Sardi sheep in Settat province, Morocco. A total of 1521 sheep, including 318 lambs < 6 months, 572 young sheep aged 6-24 months, and 631 adult sheep > 24 months, were examined from 57 flocks. Superficial CL prevalence levels were as follows: 69/318 in lambs, 292/572 in young sheep, and 114/631 in adult sheep. Additionally, 2124 sheep, including 1813 young and 311 adult sheep, were inspected in slaughterhouses ante- and post-mortem. Among all infected animals, the prevalence of visceral abscesses was notably higher in adult sheep (83%, OR = 9.8, 95% CI = 5.5-17.2) compared to young sheep (35%). Data on flock size, sex, age, body condition score, rearing methods, and hygienic measures were collected. Suspected CL cases were confirmed using quadruplex-PCR. Poor barn ventilation, lack of abscess control, and younger age were identified as key risk factors for CL. Additionally, a high percentage (89%) of animals with thoracic abscesses did not display superficial lesions. While age and abscess control have been reported in previous studies, barn ventilation appears to be a new, but important, risk factor. In conclusion, the identified risk factors in Settat province are linked to breeding management practices. Implementing regular abscess control measures and improving barn ventilation are essential for CL prevention.

Identification of hard ticks (Acari: Ixodidae) and seroprevalence to Theileria parva in cattle raised in North Kivu Province, Democratic Republic of Congo.

This study aimed to identify tick species and to determine their relationship with the Theileria parva seroprevalence in cattle raised under an extensive farming system in North Kivu Province, Democratic Republic of Congo in two agro-ecological zones namely medium (1,000-1,850 m) and high (>1,850 m) altitude. Among the 3,215 ticks collected on 482 animals, from February to April 2009, Rhipicephalus appendiculatus (64.26 %), the main vector of T. parva, was the most abundant species followed by Rhipicephalus decoloratus (35.49 %) and Amblyomma variegatum (0.25 %). The mean burden of R. appendiculatus tick per infested animal appeared significantly higher at medium (6.5 ± 0.22 ticks) than at high (0.07 ± 0.3 ticks) altitude (P < 0.05). However, an indirect fluorescent antibody test carried out on 450 blood samples revealed a global T. parva seroprevalence of 43 % (95 % CI: 38-47) which was not significantly (P > 0.05) different between medium (48.4 %; 95 % CI: 38-49) and high (41.9 %; 95 % CI: 35-49) altitude. These relatively low seroprevalences suggest that there is a state of endemicity to T. parva infection in the study area. The presence of the tick vector on animals was associated with an increased risk of being seropositive to T. parva infection (odds ratio = 2.04; 95 % CI: 1.8-2.3; P < 0.001). The results suggest the need for a longitudinal study to investigate the seasonal dynamics of tick species and T. parva infection. The rate of tick infection should also be evaluated in order to determine the intensity of T. parva transmission to cattle.

Evaluation of dog vaccination schemes against rabies in Kinshasa, Democratic Republic of the Congo.

The traditional rabies control strategy based on annual mass vaccination of dogs appears to be costly and cumbersome. Given the existence of different risk zones for rabies transmission, the present study aimed at proposing risk-based vaccination schemes by considering canine population dynamics as well as vaccine efficacy and duration of immunity (DOI). The capital of the Democratic Republic of the Congo (RDC), Kinshasa, was chosen as study site. The turnover rate of dogs was used to assess their population dynamics in two low-roaming (<25 % of dogs are roaming) and in two high-roaming zones (>75 % of dogs are roaming). The sero-conversion rate was assessed in response to primo-vaccination in three age groups: 24 puppies (≤3months), 37 juveniles (4-12 months) and 22 adult dogs. The DOI was evaluated serologically by revaccinating dogs previously vaccinated since 1-2 years (n = 31), 2-3 years (n = 12) or 3-7.5 years (n = 4). Rapid Fluorescent Focus Inhibition Test was used to quantify antibodies. These data were used to implement vaccination outcome models.The turnover rate was twice as high in high-roaming zones (36 %) as that in lowroaming zones (17 %). Irrespective of roaming level, 75 % of dogs were less than 3 years old. The vaccine was equally effective in puppies (96 %), juvenile (97 %) and adult dogs (100 %, p = 0.24). The vaccine was effective in 93 % (11/12) of puppies without pre-vaccinal protective titers (≥0.5 IU/mL). The anamnestic response was strong within 5-8 days upon the booster vaccination, in 96 % (45/47) of dogs reported vaccinated 1-7.5 years before. This suggests that the vaccine provided a long-term protection (≥3 years) which is likely to occur in 75 % of dogs in Kinshasa.Hypothesizing a vaccination stop, the vaccination outcome model allowed to estimate the time point after which vaccination coverage would drop below 40 % in function of dog population turnover rate. The systematic vaccination of puppies as well as annual vaccination of dogs aged between 3 and 15 months or annual vaccination of all unvaccinated dogs aged more than 3 months of age appeared as valuable alternative to systematic annual mass vaccination.In conclusion, this study developed a vaccination outcome model pointing out the impact of dog population dynamics and of effective duration of immunity. It appears as a promising tool for designing cost-effective rabies vaccination campaigns.

Using qualitative approaches to explore the challenges of integrated programmes for zoonosis control in developing countries: Example of hydatidosis control in Morocco.

Cystic echinococcosis, rabies and bovine tuberculosis are major neglected zoonoses in many developing countries, including Morocco. A common step in their control strategies is the management of slaughterhouses and/or the control of dog populations. This study used a qualitative approach to identify the barriers to the implementation of the cystic echinococcosis control programme in Morocco that focused on slaughterhouses and dog control. This approach could be used to understand the causes leading to the defective management of other zoonoses, and to go beyond the often-invoked 'lack of means' reason to explain failure in disease control in the Global South. Specifically, slaughterhouse observation and in-depth individual interviews with stakeholders involved in the cystic echinococcosis control programme were carried out in five Moroccan regions (Rabat-Sale-Kenitra, Mellal-Khenifra, Sous-Masaa, Laayoune-Sakia El Hamra and Guelmim-Oued Noun) for 3 years (from 2014 to 2016). Interviews (n = 81) were with agents responsible of the services managing slaughterhouses and dog population control, and also with slaughterers. Recordings were fully transcribed and narratives were analysed with the RQDA package and the R software. Triangulations and the criteria proposed by Guba and Lincon were used to assess the approach validity. This analysis revealed that in addition to 'lack of means', zoonosis control programmes can be hindered by overlapping authority/functions among the involved authorities, conflicts of interest, lack of proper training and professional practices. Zoonosis control requires the commitment of all structures, the establishment of a national and inter-regional strategy, and the updating of the legislative arsenal.

Feasibility and acceptability of a new integrated approach to control cystic echinococcosis in Morocco: Vaccination of sheep and anthelmintic treatment of dogs.

Cystic echinococcosis (CE) or hydatidosis is a common parasitic disease worldwide, especially in poor and developing countries. In Morocco, CE is a major zoonosis, despite the implementation of a national control program since 2007. Therefore, in 2016, a field trial that associates the EG95 vaccine (lambs) and anthelmintic treatment (dogs) was started in the Mid Atlas, the most endemic region in Morocco, with preliminary positive results. Here, a qualitative approach was used to analyze the feasibility and social acceptability of this strategy. Fifteen focus group discussions were separately conducted with breeders and with their wives, and forty-two individual interviews were performed with private-sector veterinarians and officers from structures responsible for the CE control program. Recordings were transcribed and analyzed with the R software, using the RQDA package. This qualitative research was validated using the credibility, transferability, dependability and confirmability criteria. This study showed that the professionals' views on sheep vaccination and anthelmintic treatment were divided between acceptability and concerns. Conversely, breeders and their wives highlighted the issue of the costs of procedures the utility of which was not immediately clear to them. All participants proposed solutions to improve this strategy, and also stressed the lack of education on CE. By bringing together the views of the communities and the professionals, this study traced the main lines (targeting the different aspects of CE and taking into account the local socio-cultural beliefs) that must be taken into account to ensure the short- and long-term CE control in Morocco.

Stakeholder analysis to improve the national control program of cystic echinococcosis in Morocco.

Cystic echinococcosis (CE) is a major zoonosis in Morocco. Despite a national hydatidosis control programme (NHCP) established in 2005, CE burden is still high nationwide. The aim of this study was to carry out an NHCP stakeholders' analysis to identify weak points and propose improvement strategies. In total, 164 semi-structured interviews were carried out with national and local stakeholder representatives, and one stakeholders' workshop was organized. Classical categories of stakeholder analysis (i.e. involvement, interest, importance, influence, priority, and power) were semi-quantified. Based on these categories, an analytical framework was proposed, by selecting elements from the stakeholder analysis, to structure the assessment around three criteria: measure appropriation, political agency, and socio-technical agency. Besides specific weak points of the NHCP, the analysis highlighted the complexity of implementing a zoonotic disease control programme due to the involvement of many different stakeholders. Finally, this study provides a simplified stakeholder analysis method that could be used to assess other health programmes targeting zoonotic diseases, in Morocco and in similar countries.

Brucellosis in wildlife in Africa: a systematic review and meta-analysis.

This study aimed to consolidate current knowledge of wildlife brucellosis in Africa and to analyse available predictors of infection. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines were followed. Information on species, test used, test results, area, rainfall, livestock and wildlife contact and year of study were extracted. This systematic review revealed 42 prevalence studies, nine disease control articles and six articles on epidemiology. Brucella abortus, Brucella melitensis, Brucella inopinata and Brucella suis were reported in wildlife. The prevalence studies revealed serological evidence of brucellosis in buffalo, antelope (positive in 14/28 species), carnivores (4/12) and other species (7/20) over the last five decades. Buffalo populations were more likely to be infected and had a higher seroprevalence than other species; the pooled seroprevalence was 13.7% (95% CI 10.3-17.3%) in buffalo, 7.1% (95% CI 1.1-15.5%) in carnivores and 2.1% (95% CI 0.1-4.9%) in antelope. Wildlife in high rainfall areas (≥ 800 mm) were more likely to be infected, and infected populations showed higher seroprevalence in high rainfall areas and in studies published after 2000. Domestic animal contact was associated with increased seroprevalence in antelope and carnivore species, but not in buffalo, supporting the hypothesis that buffalo may be a reservoir species.

Control of cystic echinococcosis in the Middle Atlas, Morocco: Field evaluation of the EG95 vaccine in sheep and cesticide treatment in dogs.

BACKGROUND: Cystic echinococcosis (CE) is an important cause of human morbidity and mortality worldwide, particularly in Morocco and other North African countries. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the potential of three strategies to reduce Echinococcus granulosus transmission: (1) 4-monthly treatment of dogs with praziquantel, (2) vaccination of sheep with the EG95 vaccine and (3) a combination of both measures. These measures were implemented during four consecutive years in different areas of the Middle Atlas Mountains in Morocco. The outcome of the interventions was assessed through hydatid cyst (viable and non-viable) counts in liver and lungs using necropsy or in vivo ultrasound examination of the liver. A total of 402 lambs were recruited for annual vaccination with the EG95 anti-E. granulosus vaccine and 395 similar lambs were selected as non-vaccinated controls. At approximately four years of age the relative risk (estimated as odds ratio) for vaccinated sheep to have viable hydatid cysts compared with non-vaccinated controls was 3% (9.37% of the vaccinated sheep were found infected while 72.82% of the controls were infected; p = 0.002). The number of viable cysts in vaccinated animals was reduced by approximately 97% (mean counts were 0.28 and 9.18 respectively; p<0.001). An average of 595 owned dogs received 4-monthly treatment during the 44 months trial, corresponding to 91% of the owned dog population. Approximately, 5% of them were examined for E. granulosus adult worms by arecoline purge or eggs in feces (confirmed by PCR). The proportion of infected dogs significantly decreased after treatment (12% versus 35%; p<0.001). Post-treatment incidence of re-infestation corresponded to a monthly risk of 4% (95% CI: 3-6%). Treatment of owned dogs on a 4-monthly basis did not reduce the level of transmission of E. granulosus to sheep, nor did it enhance the level of control generated by vaccination of sheep with EG95, possibly because of unowned dogs and wild canids were not treated. CONCLUSIONS/SIGNIFICANCE: These data suggest that vaccination of sheep with EG95 has the potential to reduce the level of CE in Morocco and in other parts of the world with similar transmission dynamics. Under the epidemiological circumstances existing in the trial area, 4-monthly treatment of owned dogs with praziquantel was insufficient to have a major impact of E. granulosus transmission to sheep.

A cross-sectional study on bovine tuberculosis in Hawassa town and its surroundings, Southern Ethiopia.

A cross-sectional study was conducted in Hawassa town and its surroundings from October 2007 to May 2008 to estimate the prevalence of bovine tuberculosis (BTB) based on comparative interadermal tuberculin test (CIDT) and abattoir survey. Accordingly, 39 herds comprising 413 cattle were subjected to CIDT, and the herd and individual animal prevalence were 48.7% (19/39) and 11.6% (48/413), respectively. One of the 16 milk samples collected from tuberculin-positive cows was culture positive. The prevalence significantly differed among the age group (P = 0.001) and management system (P = 0.001). Thus, age group over four (OR = 7.9) and animal with poor management system (OR = 4.1) had a higher odds for tuberculin reactivity compared to those with age group under four and cattle with good management system, respectively. Of the total 1,023 cattle subjected to postmortem examination, 11 (1.1%) were found to be positive for gross tuberculous lesions. Larger proportion (50%) of TB lesion was recorded in the respiratory pathway followed by digestive system (28.6%) and prescapular lymph nodes (21.4%). Of 14 tissue specimens collected from the gross lesions, four (28.6%) were positive for histopathological TB lesions. In conclusion, this study revealed the importance of BTB in the study area in particular and the region in general.

An alternative cold chain for storing and transporting East Coast fever vaccine.

East Coast fever (ECF) is an often fatal, economically important cattle disease that predominantly affects eastern, central, and southern Africa. ECF is controlled through vaccination by means of simultaneous injection of oxytetracycline and cryogenically preserved stabilate containing live, disease-causing parasites. Storage and transportation of the stabilate requires liquid nitrogen, a commodity that is commonly unreliable in low-resource settings. Here we show that storage of conventionally prepared stabilate at -80 °C for up to 30 days does not significantly affect its ability to infect cultured peripheral blood mononucleated cells or live cattle, suggesting an alternative cold chain that maintains these temperatures could be used to effectively manage ECF.

Cystic Echinococcosis in Three Locations in the Middle Atlas, Morocco: Estimation of the Infection Rate in the Dog Reservoir.

A longitudinal study was carried out in Middle atlas, Morocco (locality of Had Oued Ifrane) in a population of 255 dogs from three localities, including two categories of dogs (owned and stray dogs). The dogs were investigated three times over a period ranging from 4 to 8 months between December and August. At each investigation, dogs were treated with arecoline, inducing defecation and allowing feces collection. Dogs were further treated with praziquantel to clear them from Echinococcus granulosus. Microscopic examination of feces was performed to assess the infection status of dogs at each investigation, and positive samples underwent copro-PCR to determine the circulating strain of E. granulosus. A high prevalence of infestation ranging from 23.5% to 38.8% and from 51.3% to 68.5% was, respectively, found in owned and in stray dogs. The PCR results revealed the presence of G1 strain in all positive samples. A logistic regression model was used to determine the incidence of infestation and showed that stray dogs underwent a significantly higher risk of infection (odds ratio = 14; 95% confidence interval: 6-30; p < 0.001) compared with owned dogs. Only anthelmintic treatment intervals of 2 months efficiently prevented egg shedding in owned and stray dogs. The seasonal effect was also significant, with the highest risk of reinfestation in winter and the lowest risk in summer. This study confirms that stray dogs undergo an increased risk of infestation by E. granulosus and indicate that infective pressure is influenced by season.

The socio-economic burden of cystic echinococcosis in Morocco: A combination of estimation method.

Cystic echinococcosis (CE) is a major zoonosis in Morocco despite the launch of a national control programme in 2005. As its economic consequences have not been studied yet in Morocco, this study estimated CE impact in terms of monetary losses, disability-adjusted life years (DALY), and DALY for zoonotic diseases (zDALY) in the entire country and in specific regions for the 2011 to 2014 period. The direct monetary losses were related to organ seizure from infected animal in slaughterhouses, and to healthcare expenses as well as lost wages for infected humans. Animal production losses concerned milk yield, fertility, carcass weight, and wool production. Losses due to human infection were also composed of disability and productivity losses at work. Monte Carlo simulations were used to estimate monetary losses and zDALY values. Nationwide, the estimated DALY was 0.5 years per 100,000 persons per year, and the zDALY was 55 years per 100,000 persons per year. Total yearly losses were estimated at 73 million USD (54-92 million USD). However, losses differed significantly among regions. Most of the economic losses consisted of unperceived consequences, i.e. decreased animal production and reduced productivity of asymptomatic individuals. Future studies should determine the socioeconomic and epidemiological factors underlying the differences in economic losses among regions to develop better adapted control programmes.

Prevalence and antimicrobial resistance of Salmonella isolated from broiler farms, chicken carcasses, and street-vended restaurants in Casamance, Senegal.

This study was undertaken to determine the prevalence and distribution of Salmonella on 57 randomly selected broiler farms at the end of the rearing period and in chicken products in urban and periurban areas in Casamance, Senegal, and to evaluate the antimicrobial resistance profiles of the Salmonella serovars. Salmonella was detected in chicken feces, on carcass skin, and in muscle on 35.1, 38.6, and 29.8% of farms, respectively. Salmonella was found in chicken meat servings from 14.3% of the 42 street restaurants and in 40.4% of the 285 chicken carcasses examined. The prevalence on skin and in muscle was significantly associated with the detection of Salmonella in feces (P

Genetic and antigenic variation of the bovine tick-borne pathogen Theileria parva in the Great Lakes region of Central Africa.

BACKGROUND: Theileria parva causes East Coast fever (ECF), one of the most economically important tick-borne diseases of cattle in sub-Saharan Africa. A live immunisation approach using the infection and treatment method (ITM) provides a strong long-term strain-restricted immunity. However, it typically induces a tick-transmissible carrier state in cattle and may lead to spread of antigenically distinct parasites. Thus, understanding the genetic composition of T. parva is needed prior to the use of the ITM vaccine in new areas. This study examined the sequence diversity and the evolutionary and biogeographical dynamics of T. parva within the African Great Lakes region to better understand the epidemiology of ECF and to assure vaccine safety. Genetic analyses were performed using sequences of two antigen-coding genes, Tp1 and Tp2, generated among 119 T. parva samples collected from cattle in four agro-ecological zones of DRC and Burundi. RESULTS: The results provided evidence of nucleotide and amino acid polymorphisms in both antigens, resulting in 11 and 10 distinct nucleotide alleles, that predicted 6 and 9 protein variants in Tp1 and Tp2, respectively. Theileria parva samples showed high variation within populations and a moderate biogeographical sub-structuring due to the widespread major genotypes. The diversity was greater in samples from lowlands and midlands areas compared to those from highlands and other African countries. The evolutionary dynamics modelling revealed a signal of selective evolution which was not preferentially detected within the epitope-coding regions, suggesting that the observed polymorphism could be more related to gene flow rather than recent host immune-based selection. Most alleles isolated in the Great Lakes region were closely related to the components of the trivalent Muguga vaccine. CONCLUSIONS: Our findings suggest that the extensive sequence diversity of T. parva and its biogeographical distribution mainly depend on host migration and agro-ecological conditions driving tick population dynamics. Such patterns are likely to contribute to the epidemic and unstable endemic situations of ECF in the region. However, the fact that ubiquitous alleles are genetically similar to the components of the Muguga vaccine together with the limited geographical clustering may justify testing the existing trivalent vaccine for cross-immunity in the region.

A significant increase in kdr in Anopheles gambiae is associated with an intensive vector control intervention in Burundi highlands.

OBJECTIVES AND METHODS: In Burundi, the occurrence of the knock down resistance (kdr) mutation in Anopheles gambiae sensu lato (s.l.) was determined for six consecutive years within the framework of a vector control programme. Findings were also linked with the insecticide resistance status observed with bioassay in An. gambiae s.l. and An. funestus. RESULTS: The proportion of An. gambiae s.l. carrying the East Leu-Ser kdr mutation was 1% before the spraying intervention in 2002; by 2007 it was 86% in sprayed valleys and 67% in untreated valleys. Multivariate analysis showed that increased risk of carrying the kdr mutation is associated with spraying interventions, location and time. In bioassays conducted between 2005 and 2007 at five sites, An. funestus was susceptible to permethrin, deltamethrin and DDT. Anopheles gambiae s.l. remained susceptible or tolerant to deltamethrin and resistant to DDT and permethrin, but only when kdr allele carriers reached 90% of the population. CONCLUSIONS: The cross-resistance against DDT and permethrin in Karuzi suggests a possible kdr resistance mechanism. Nevertheless, the homozygous resistant genotype alone does not entirely explain the bioassay results, and other mechanisms conferring resistance cannot be ruled out. After exposure to all three insecticides, homozygote individuals for the kdr allele dominate among the surviving An. gambiae s.l. This confirms the potential selection pressure of pyrethroids on kdr mutation. However, the high occurrence of the kdr mutation, even at sites far from the sprayed areas, suggests a selection pressure other than that exerted by the vector control programme.

The sensitivity of PCR and serology in different Theileria parva epidemiological situations in Rwanda.

Theileria parva is the causative agent of a lethal tick-borne disease of cattle occurring in eastern, central and southern Africa. Variations in the sensitivity of the serological and molecular tests with seasonal vector occurrence and discrepancies between low PCR prevalence and high T. parva vector density are a setback to estimate true prevalences. Therefore, the objectives of the present studies were to evaluate (1) the sensitivity of three serological tests (IFAT, ELISA and SELISA) and one molecular test (PCR) in the diagnosis of chronic T. parva infections in four different agro-ecological zones of Rwanda and (2) the effect of tick challenge and animal's age on the sensitivity of PCR. Blood samples from 635 bovines were collected in four agro-ecological zones of Rwanda. All sera were screened using the IFAT, ELISA, SELISA and PCR. The binary results of the four diagnostic tests were introduced separately for each agro-ecological zone in a Bayesian model to estimate the prevalence of T. parva infections and the sensitivity of the four diagnostic tests. All test specificities were set to 100%. The estimated T. parva prevalence was much higher (83-85%) than estimations based on single diagnostic tests. The estimated sensitivity of serological tests was relatively constant and ranged from 57 to 75% in the various areas. The sensitivity of PCR showed more pronounced variations, ranging from 66% in the low T. parva transmission (high land) zones compared to 24% in the highly vector infested (low land) zones. Calves and adult cattle (n=194) were also sampled in regularly and irregularly dipped herds in the low land region. The apparent T. parva prevalence detected by PCR was significantly higher in calves than in adult cattle and in herds regularly treated with acaricides, while no significant differences were found with IFAT. The conditional probability that a sample was positive at PCR while it was positive at IFAT was significantly lower in adults. The implication of these findings in the use of diagnostic assays for epidemiological studies is discussed.

Immunological response to Brucella abortus strain 19 vaccination of cattle in a communal area in South Africa.

Brucellosis is of worldwide economic and public health importance. Heifer vaccination with live attenuated Brucella abortus strain 19 (S19) is the cornerstone of control in low- and middle-income countries. Antibody persistence induced by S19 is directly correlated with the number of colony-forming units (CFU) per dose. There are two vaccination methods: a 'high' dose (5-8 × 1010 CFU) subcutaneously injected or one or two 'low' doses (5 × 109 CFU) through the conjunctival route. This study aimed to evaluate serological reactions to the 'high' dose and possible implications of the serological findings on disease control. This study included 58 female cases, vaccinated at Day 0, and 29 male controls. Serum was drawn repeatedly and tested for Brucella antibodies using the Rose Bengal Test (RBT) and an indirect enzyme-linked immunosorbent assay (iELISA). The cases showed a rapid antibody response with peak RBT positivity (98%) at 2 weeks and iELISA (95%) at 8 weeks, then decreased in an inverse logistic curve to 14% RBT and 32% iELISA positive at 59 weeks and at 4.5 years 57% (4/7 cases) demonstrated a persistent immune response (RBT, iELISA or Brucellin skin test) to Brucella spp. Our study is the first of its kind documenting the persistence of antibodies in an African communal farming setting for over a year to years after 'high' dose S19 vaccination, which can be difficult to differentiate from a response to infection with wild-type B. abortus. A recommendation could be using a 'low' dose or different route of vaccination.