Genomic epidemiology of carbapenemase-producing Gram-negative bacteria at the human-animal-environment interface in Djibouti city, Djibouti.

The emergence of carbapenem resistance is a major public health threat in sub-Saharan Africa but remains poorly understood, particularly at the human-animal-environment interface. This study provides the first One Health-based study on the epidemiology of Carbapenemase-Producing Gram-Negative Bacteria (CP-GNB) in Djibouti City, Djibouti, East Africa. In total, 800 community urine samples and 500 hospital specimens from humans, 270 livestock fecal samples, 60 fish samples, and 20 water samples were collected and tested for carbapenem resistance. The overall estimated CP-GNB prevalence was 1.9 % (32/1650 samples) and specifically concerned 0.3 % of community urine samples, 2.8 % of clinical specimens, 2.6 % of livestock fecal samples, 11.7 % of fish samples, and 10 % of water samples. The 32 CP-GNB included 19 Escherichia coli, seven Acinetobacter baumannii, five Klebsiella pneumoniae, and one Proteus mirabilis isolate. Short-read (Illumina) and long-read (Nanopore) genome sequencing revealed that carbapenem resistance was mainly associated with chromosomal carriage of blaNDM-1, blaOXA-23, blaOXA-48, blaOXA-66, and blaOXA-69 in A. baumannii, and with plasmid carriage in Enterobacterales (blaNDM-1 and blaOXA-181 in E. coli, blaNDM-1, blaNDM-5 and blaOXA-48 in K. pneumoniae, and blaNDM-1 in P. mirabilis). Moreover, 17/32 CP-GNB isolates belonged to three epidemic clones: (1) A. baumannii sequence type (ST) 1697,2535 that showed a distribution pattern consistent with intra- and inter-hospital dissemination; (2) E. coli ST10 that circulated at the human-animal-environment interface; and (3) K. pneumoniae ST147 that circulated at the human-environment interface. Horizontal exchanges probably contributed to carbapenem resistance dissemination in the city, especially the blaOXA-181-carrying ColKP3-IncX3 hybrid plasmid that was found in E. coli isolates belonging to different STs. Our study highlights that despite a relatively low CP-GNB prevalence in Djibouti City, plasmids harboring carbapenem resistance circulate in humans, animals and environment. Our findings stress the need to implement preventive and control measures for reducing the circulation of this potentially emerging public health threat.

Seroprevalence of SARS-CoV-2 IgG and associated factors among people living with HIV over the first 12 months following the outbreak of COVID-19 in Burkina Faso, a sub-Saharan African country.

OBJECTIVE: This study aimed to evaluate the seroprevalence of anti-SARS-CoV-2 IgG and factors associated with the infection among PLWHIV over the first 12 months following the outbreak of COVID-19 in Burkina Faso. DESIGN: A retrospective cross-sectional study of plasma samples collected from March 9, 2020, and March 8, 2021, at the outpatient HIV referral center, before the introduction of the SARS-CoV-2 vaccine in Burkina Faso. METHODS: Anti-SARS-CoV-2 IgG were detected in plasma using DS-ЕIA-ANTI-SARS-CoV-2-G (S) kit. Logistic regressions were used to compare SARS-CoV-2 specific immune responses between groups and within subgroups. RESULTS AND DISCUSSION: A total of 419 plasma were subjected to serological diagnosis. None of the participants was vaccinated against COVID-19 during the period of sample collection, and 130 samples were positive for anti-SARS-CoV-2 IgG, giving a prevalence of 31.0% (95% CI 26.6-35.7). The median CD4 cell count was 661 cells/µL (IQR,422-928). Retailers had half the risk of being infected compared to housemaids with an OR of 0.49 (p = 0.028, 95% CI 0.26-0.91). Likewise, the risk of infection was 1.69 times higher in patients on integrase inhibitors compared to that of patients on non-nucleoside reverse transcriptase inhibitors (p = 0.020, 95% CI 1.09-2.63). CONCLUSION: Our study reveals a high seroprevalence among PLWHIV to SARS-CoV-2 during the first year of the pandemic. In addition, PLWHIV on integrase inhibitors are 1.69 times more likely to be infected than PLWHIV on non-nucleoside inhibitors, and this observation remains an intriguing topic that still needs to be clarified.

Genetic Diversity and Population Structure of Mycobacterium bovis at the Human-Animal-Ecosystem Interface in France: "A One Health Approach".

Mycobacterium bovis infects cattle and wildlife, and also causes a small proportion of tuberculosis cases in humans. In most European countries, M. bovis infections in cattle have been drastically reduced, but not eradicated. Here, to determine the M. bovis circulation within and between the human, cattle, and wildlife compartments, we characterized by spoligotyping and mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing the genetic diversity of M. bovis isolates collected from humans, cattle, and wildlife in France from 2000 to 2010. We also assessed their genetic structure within and among the different host groups, and across time and space. The M. bovis genetic structure and its spatiotemporal variations showed different dynamics in the human and animal compartments. Most genotypes detected in human isolates were absent in cattle and wildlife isolates, possibly because in patients, M. bovis infection was contracted abroad or was the reactivation of an old lesion. Therefore, they did not match the genetic pool present in France during the study period. However, some human-cattle exchanges occurred because some genotypes were common to both compartments. This study provides new elements for understanding M. bovis epidemiology in France, and calls for increased efforts to control this pathogen worldwide.

Seroprevalence and risk factors of Toxoplasma gondii infection in sheep in Algeria.

Toxoplasma gondii infection is recognized as one of the major causes of reproductive failure in sheep and goats. This survey was carried out in order to study the seroprevalence of Toxoplasma infection in sheep in Blida, Bouira and Medea regions from Algeria. The sample size was set at 220 animals distributed over 22 farms. Sera were assayed for T. gondii antibody detection by Modified Agglutination Test (MAT). The overall seroprevalence was 35.9% (79/220) with a herd seroprevalence of 77.3% (17/22). The prevalence was significantly higher in Medea (45.7% of 116 sheep), compared to Blida (27.7% of 83 sheep). Bouira region showed the lowest prevalence with 3 positive samples (14.3%) over 21 sheep. Logistic regression analysis revealed that the likelihood of T. gondii infection was higher in semi-extensive sheep breeding, in regions where the presence of cats is strong, and in highlands when compared with semi-intensive sheep breeding, weak presence of cat and in lowland, respectively. This study shows a high seroprevalence of Toxoplasma infection in sheep in these areas.

Multidrug-Resistant Enterobacterales in Community-Acquired Urinary Tract Infections in Djibouti, Republic of Djibouti.

The emergence and spread of multidrug resistant Enterobacterales (MDR-E) are a global public health issue. This problem also concerns urinary tract infections (UTI), which are the second most frequent infections after respiratory infections. The objective of this study was to determine MDR-E frequency and to characterize MDR-E isolates from patients with community-acquired UTIs in Djibouti, Republic of Djibouti. From 800 clinical urinary samples collected at the Mer Rouge Laboratory, Djibouti, from January to July 2019, 142 were identified as Enterobacterales (age range of the 142 patients mean age is 42 years.) Mass spectrometry analysis of these isolates identified 117 Escherichia coli, 14 Klebsiella pneumoniae, 2 Proteus mirabilis, 4 Enterobacter spp., 4 Providencia stuartii and 1 Franconibacter helveticus. Antibiotic susceptibility testing (disk diffusion method) of these 142 isolates detected 68 MDR-E (68/142 = 48%): 65 extended-spectrum bêta lactamase- (ESBL), 2 carbapenemase- (one also ESBL), and 1 cephalosporinase-producer. Multiplex PCR and sequencing showed that the 65 ESBL-producing isolates carried genes encoding CTX-M enzymes (CTX-M-15 in 97% and CTX-M-9 in 3% of isolates). Two isolates harboured a gene encoding the OXA-48-like carbapenemase, and one the gene encoding the AmpC CMY-2 cephalosporinase. Genes implicated in resistance to quinolones (qnrB, aac (6')-Ib-cr, qnrD, oqxA and B) also were detected. Among the E. coli phylogroups, B2 was the most common phylogenetic group (21% of MDR-E isolates and 26% of non-MDR-E isolates), followed by A (14% and 12%), B1 (9% and 7%), D (3% and 3%), F (3% and 3%) and E (2% and 2%). This study highlights the high frequency of ESBL producers and the emergence of carbapenemase-producers among Enterobacterales causing community-acquired UTIs in Djibouti.

Genomic Epidemiology of SARS-CoV-2 in Western Burkina Faso, West Africa.

BACKGROUND: After its initial detection in Wuhan, China, in December 2019, SARS-CoV-2 has spread rapidly, causing successive epidemic waves worldwide. This study aims to provide a genomic epidemiology of SARS-CoV-2 in Burkina Faso. METHODS: Three hundred and seventy-seven SARS-CoV-2 genomes obtained from PCR-positive nasopharyngeal samples (PCR cycle threshold score < 35) collected between 5 May 2020, and 31 January 2022 were analyzed. Genomic sequences were assigned to phylogenetic clades using NextClade and to Pango lineages using pangolin. Phylogenetic and phylogeographic analyses were performed to determine the geographical sources and time of virus introduction in Burkina Faso. RESULTS: The analyzed SARS-CoV-2 genomes can be assigned to 10 phylogenetic clades and 27 Pango lineages already described worldwide. Our analyses revealed the important role of cross-border human mobility in the successive SARS-CoV-2 introductions in Burkina Faso from neighboring countries. CONCLUSIONS: This study provides additional insights into the genomic epidemiology of SARS-CoV-2 in West Africa. It highlights the importance of land travel in the spread of the virus and the need to rapidly implement preventive policies. Regional cross-border collaborations and the adherence of the general population to government policies are key to prevent new epidemic waves.

Molecular prevalence, genetic characterization and patterns of Toxoplasma gondii infection in domestic small mammals from Cotonou, Benin.

Toxoplasmosis, one of the most prevalent parasitic infections in humans and animals, is caused by the intracellular protozoan parasite Toxoplasma gondii. Small mammals play a key role as intermediate reservoir hosts in the maintenance of the T. gondii life cycle. In this study, we estimated the molecular prevalence and provide genetic diversity data for T. gondii in 632 small mammals sampled in four areas of Cotonou city, Benin. Both the brain and heart of each individual were screened through T. gondii-targeting qPCR, and positive samples were then genotyped using a set of 15 T. gondii-specific microsatellites. Prevalence data were statistically analyzed in order to assess the relative impact of individual host characteristics, spatial distribution, composition of small mammal community, and urban landscape features. An overall T. gondii molecular prevalence of 15.2% was found and seven genotypes, all belonging to the Africa 1 lineage, could be retrieved from the invasive black rat Rattus rattus and the native African giant shrew Crocidura olivieri. Statistical analyses did not suggest any significant influence of the environmental parameters used in this study. Rather, depending on the local context, T. gondii prevalence appeared to be associated either with black rat, shrew, or mouse abundance or with the trapping period. Overall, our results highlight the intricate relationships between biotic and abiotic factors involved in T. gondii epidemiology and suggest that R. rattus and C. olivieri are two competent reservoirs for the Africa 1 lineage, a widespread lineage in tropical Africa and the predominant lineage in Benin.

Title: Prévalence moléculaire, caractérisation génétique et schémas d'infection par Toxoplasma gondii chez les petits mammifères domestiques de Cotonou, Bénin. Abstract: La toxoplasmose, l'une des infections parasitaires les plus répandues chez l'homme et les animaux, est causée par le parasite protozoaire intracellulaire Toxoplasma gondii. Les petits mammifères jouent un rôle clé en tant qu'hôtes réservoirs intermédiaires dans le maintien du cycle de vie de T. gondii. Dans cette étude, nous estimons sa prévalence moléculaire et fournissons des données sur sa diversité génétique chez 632 petits mammifères échantillonnés dans quatre localités de la ville de Cotonou. Le cerveau et le cœur de chaque individu ont été analysés par qPCR ciblant T. gondii, et les échantillons positifs ont ensuite été génotypés à l'aide d'un ensemble de 15 microsatellites spécifiques à T. gondii. Les données de prévalence ont été analysées statistiquement afin d'évaluer l'impact relatif des caractéristiques individuelles de l'hôte, de la distribution spatiale, de la composition de la communauté des petits mammifères ainsi que des caractéristiques du paysage urbain. Une prévalence moléculaire globale de T. gondii de 15,2 % a été estimée et sept génotypes, tous appartenant à la lignée Africa 1, ont pu être extraits du rat noir Rattus rattus, espèce envahissante, et de la musaraigne Crocidura olivieri, espèce indigène. Les analyses statistiques n'ont pas suggéré d'influence significative des paramètres environnementaux utilisés dans cette étude. Au contraire, selon le contexte local, la prévalence de T. gondii semble être associée à l'abondance de rats noirs, de musaraignes ou de souris ainsi qu'à la période de piégeage. Dans l'ensemble, nos résultats mettent en évidence les relations complexes entre les facteurs biotiques et abiotiques impliqués dans l'épidémiologie de T. gondii et suggèrent que R. rattus et C. olivieri sont deux réservoirs compétents pour la lignée Africa 1, une lignée répandue en Afrique tropicale et prédominante au Bénin.

A unique Toxoplasma gondii haplotype accompanied the global expansion of cats.

Toxoplasma gondii is a cyst-forming apicomplexan parasite of virtually all warm-blooded species, with all true cats (Felidae) as definitive hosts. It is the etiologic agent of toxoplasmosis, a disease causing substantial public health burden worldwide. Few intercontinental clonal lineages represent the large majority of isolates worldwide. Little is known about the evolutionary forces driving the success of these lineages, the timing and the mechanisms of their global dispersal. In this study, we analyse a set of 156 genomes and we provide estimates of T. gondii mutation rate and generation time. We elucidate how the evolution of T. gondii populations is intimately linked to the major events that have punctuated the recent history of cats. We show that a unique haplotype, whose length represents only 0.16% of the whole T. gondii genome, is common to all intercontinental lineages and hybrid populations derived from these lineages. This haplotype has accompanied wildcats (Felis silvestris) during their emergence from the wild to domestic settlements, their dispersal in the Old World, and their expansion in the last five centuries to the Americas. The selection of this haplotype is most parsimoniously explained by its role in sexual reproduction of T. gondii in domestic cats.

Toxoplasma gondii in the faeces of wild felids from the Atlantic Forest, Brazil.

BACKGROUND: Toxoplasma gondii is a apicomplexan parasite of virtually all warm-blooded species. All true cats (Felidae) can act as definitive hosts for this parasite by shedding resistant oocysts into the environment. However, the patterns of oocysts shedding are only partially understood in domestic cats and largely unknown in wild felids. OBJECTIVES: We carried out molecular analysis of 82 faecal samples from wild felids collected in the Serra dos Órgãos National Park (Parnaso), Rio de Janeiro, Brazil. METHODS: We screened samples for T. gondii DNA using a quantitative polymerase chain reaction (qPCR) targeting the 529bp DNA fragment. Polymerase chain reaction (PCR)-positive samples were genotyped using 15 microsatellite markers. RESULTS: Only one faecal sample from a Puma yagouaroundi was PCR-positive [cycle threshold (Ct) = 26.88]. This sample was contaminated by a T. gondii strain of BrIII lineage, a common lineage in domestic animals from Brazil. MAIN CONCLUSIONS: This first report of T. gondii in faeces of wild South American felids in their natural environment indicates infrequent oocyst shedding and suggests a role of acquired immunity in limiting re-excretion as in domestic cats. The presence of a domestic strain of T. gondii in a faecal sample from a wild felid at very low concentrations (not detected by microscopy) is consistent with the hypothesis of host-parasite co-adaptations limiting the circulation of T. gondii strains between domestic and wild environments.

Toxoplasma gondii in the faeces of wild felids from the Atlantic Forest, Brazil

BACKGROUND Toxoplasma gondii is a apicomplexan parasite of virtually all warm-blooded species. All true cats (Felidae) can act as definitive hosts for this parasite by shedding resistant oocysts into the environment. However, the patterns of oocysts shedding are only partially understood in domestic cats and largely unknown in wild felids. OBJECTIVES We carried out molecular analysis of 82 faecal samples from wild felids collected in the Serra dos Órgãos National Park (Parnaso), Rio de Janeiro, Brazil. METHODS We screened samples for T. gondii DNA using a quantitative polymerase chain reaction (qPCR) targeting the 529bp DNA fragment. Polymerase chain reaction (PCR)-positive samples were genotyped using 15 microsatellite markers. RESULTS Only one faecal sample from a Puma yagouaroundi was PCR-positive [cycle threshold (Ct) = 26.88]. This sample was contaminated by a T. gondii strain of BrIII lineage, a common lineage in domestic animals from Brazil. MAIN CONCLUSIONS This first report of T. gondii in faeces of wild South American felids in their natural environment indicates infrequent oocyst shedding and suggests a role of acquired immunity in limiting re-excretion as in domestic cats. The presence of a domestic strain of T. gondii in a faecal sample from a wild felid at very low concentrations (not detected by microscopy) is consistent with the hypothesis of host-parasite co-adaptations limiting the circulation of T. gondii strains between domestic and wild environments.

First isolation and genotyping of Toxoplasma gondii strains from domestic animals in Tunisia.

The isolation and molecular typing of Toxoplasma gondii strains provide an essential basis for a better understanding of the parasite's genetic diversity, determinants of its geographical distribution and associated risks to human health. In this study, we isolated and genetically characterized T. gondii strains from domestic animals in Southern and coastal area of Tunisia. Blood, hearts and/or brains were collected from 766 domestic animals (630 sheep and 136 free-range chickens). Strain isolation from these samples was performed using mouse bioassay and genotyping was carried out with a multiplex PCR technique using 15 microsatellite markers. Thirty viable strains of T. gondii were successfully isolated from tissues of sheep (19/142) and chickens (11/33). In addition, 3 strains could be successfully genotyped from animal tissues for which mouse bioassay was unsuccessful. A large predominance of type II strains (n = 29) was found in the sampled regions, followed by type III (n = 3) and, for the first time in Tunisia, a single isolate of Africa 4 lineage from a sheep. Analyses of population genetics showed the presence of a divergent population of type II lineage in Tunisia, supporting limited recent migrations of strains between Tunisia and other countries of the world.

A hotspot of Toxoplasma gondii Africa 1 lineage in Benin: How new genotypes from West Africa contribute to understand the parasite genetic diversity worldwide.

Through international trades, Europe, Africa and South America share a long history of exchanges, potentially of pathogens. We used the worldwide parasite Toxoplasma gondii to test the hypothesis of a historical influence on pathogen genetic diversity in Benin, a West African country with a longstanding sea trade history. In Africa, T. gondii spatial structure is still non-uniformly studied and very few articles have reported strain genetic diversity in fauna and clinical forms of human toxoplasmosis so far, even in African diaspora. Sera from 758 domestic animals (mainly poultry) in two coastal areas (Cotonou and Ouidah) and two inland areas (Parakou and Natitingou) were tested for T. gondii antibodies using a Modified Agglutination Test (MAT). The hearts and brains of 69 seropositive animals were collected for parasite isolation in a mouse bioassay. Forty-five strains were obtained and 39 genotypes could be described via 15-microsatellite genotyping, with a predominance of the autochthonous African lineage Africa 1 (36/39). The remaining genotypes were Africa 4 variant TUB2 (1/39) and two identical isolates (clone) of Type III (2/39). No difference in terms of genotype distribution between inland and coastal sampling sites was found. In particular, contrarily to what has been described in Senegal, no type II (mostly present in Europe) was isolated in poultry from coastal cities. This result seems to refute a possible role of European maritime trade in Benin despite it was one of the most important hubs during the slave trade period. However, the presence of the Africa 1 genotype in Brazil, predominant in Benin, and genetic analyses suggest that the triangular trade was a route for the intercontinental dissemination of genetic strains from Africa to South America. This supports the possibility of contamination in humans and animals with potentially imported virulent strains.

Seroprevalence of Toxoplasma gondii among healthy blood donors in two locations in Tunisia and associated risk factors.

Toxoplasma gondii is a protozoan parasite that can be transmitted to humans through a variety of routes including blood transfusion. This study aimed to investigate the seroprevalence of T. gondii infection and associated risk factors in healthy blood donors in Tunisia. A total of 800 healthy blood donors from two blood centers in south and coastal Tunisia were analyzed for anti-T. gondii IgG and IgM antibodies by indirect immunofluorescence assay (IFA) and enzyme-linked immunoassays (ELISA), respectively. Structured questionnaires were used to gather information on risk factors for T. gondii infection during collection. The overall seroprevalence was 44.4% of which 352 (44%) and 3 (0.4%) were positive for IgG and both IgG and IgM anti-T. gondii antibodies, respectively. Multivariate analysis showed that T. gondii seropositivity was significantly associated with the birth place (adjusted odds ratio [OR] = 2.72; 95% confidence interval [CI]: 1.49-4.94) and the age of the donors (adjusted OR = 4.98; 95% CI: 1.50-16.58) which are independent risk factors. In addition, the variables of hand washing before eating (adjusted OR = 0.52; 95% CI: 0.37-0.74) and living in an urban environment (adjusted OR = 0.30; 95% CI: 0.13-0.71) are two protective factors. This study provided the first data on the seroprevalence and epidemiology of T. gondii infection in healthy blood donors in Tunisia.

TITLE: Séroprévalence de Toxoplasma gondii chez des donneurs de sang sains dans deux sites en Tunisie et facteurs de risque associés. ABSTRACT: Toxoplasma gondii est un parasite protozoaire qui peut être transmis à l'homme par diverses voies, dont la transfusion sanguine. Cette étude vise à étudier la séroprévalence de l'infection à T. gondii et les facteurs de risque associés chez les donneurs de sang sains en Tunisie. Au total, huit cents donneurs de sang sains de deux centres de transfusion sanguine du sud et de la côte tunisienne ont été analysés respectivement pour la recherche des anticorps IgG et IgM anti-T. gondii par immunofluorescence indirecte (IFA) et par dosage immuno-enzymatique (ELISA). Des questionnaires structurés ont été utilisés pour recueillir des informations sur les facteurs de risque d'infection à T. gondii pendant la collecte. La séroprévalence globale était de 44,4 % dont 352 (44 %) et 3 (0,4 %) étaient respectivement positifs pour les anticorps IgG et IgG/IgM anti-T. gondii. Une analyse multivariée a montré que la séropositivité à T. gondii était significativement associée au lieu de naissance (rapport de côtes ajusté [OR] = 2,72 ; intervalle de confiance à 95 % [IC] : 1,49­4,94) et à l'âge des donneurs (OR ajusté = 4,98 ; IC 95 % : 1,50­16,58) qui sont des facteurs de risque indépendants. De plus, le lavage des mains avant de manger (OR ajusté = 0,52 ; IC 95 % : 0,37­0,74) et vivre dans un milieu urbain (OR ajusté = 0,30 ; IC 95 % : 0,13­0,71) sont deux facteurs de protection. Cette étude a fourni les premières données sur la séroprévalence et l'épidémiologie de l'infection à T. gondii chez les donneurs de sang sains en Tunisie.

Seroprevalence and Risk Factors of Toxoplasma gondii Infection in Free-Range Chickens in Senegal, West Africa.

In sub-Saharan Africa, few studies have addressed the environmental determinants of the incidence of Toxoplasma gondii infection. Free-range chickens are regarded as sensitive indicators for environmental contamination with T. gondii oocysts due to their ground-feeding behavior and have been used as sentinels. A cross-sectional study was conducted from January to April 2018 to estimate the seroprevalence of T. gondii infection in free-range chickens from Senegal, West Africa, using the modified agglutination test. Sampling was performed in two regions of the country: Saint-Louis, a Sahelian area in the North, and Kedougou, a forest and rainy area in the South. A questionnaire was administered to chicken owners to assess potential risk factors for T. gondii seropositivity, and univariable and multivariable logistic regression analyses were used to determine the statistical significance of risk factors. The seroprevalence in chickens was 7.67% (51/665; 95% confidence interval [CI]: 5.51-9.52). The multivariable logistic regression model indicated that the odds to test T. gondii seropositive was higher in chickens provided well water (odds ratio [OR] = 3.17, 95% CI: 1.45-6.93; p = 0.004) than in chickens provided tap water, and in hens having chicks (OR = 80.15, 95% CI: 22.79-281.95; p < 0.001) than in chickens (male or female) not having chicks. The possible role of contaminated well water in the acquisition of infection in chickens-and hence in human-merits consideration and should be addressed in future studies.

Combining spatial analysis and host population genetics to gain insights into the mode of transmission of a pathogen: The example of Toxoplasma gondii in mice.

Toxoplasma gondii is a ubiquitous highly prevalent zoonotic protozoan. Cats are the definitive hosts, while all other warm-blooded animals are intermediate hosts for this parasite. Commensal rodents, being the main prey of cats, are probably the major reservoir for T. gondii in the domestic environment. Rodents can acquire infection after ingestion of oocysts that have sporulated in the environment. However, experimental evidence shows that vertical transmission can be sufficient for the perpetuation of transmission between generations of mice. In natural settings, the relative epidemiological importance of vertical transmission over oral transmission is a matter of debate and raises the question of the possibility of a T. gondii cycle in the absence of cats. In the present study, we took advantage of an extensive survey of commensal rodents in Dakar, Senegal, where the house mouse is the predominant putative reservoir of T. gondii. Mice genotypes and spatial location through GPS referencing of all trapping localizations were investigated in relation to T. gondii infection in eight sites of the city of Dakar and on Goree Island. In each sampling site, the occurrence of over-prevalence zones of T. gondii infection was investigated through Kulldorf's statistic using SaTScan software. Genetic structure and relatedness between mice were investigated within each over-prevalence zone, in order to find clues of transmission between related mice. Within each of the four over-prevalence zones identified across nine sites, infected mice belonged to more than one genetic group. No association between the degree of relatedness and the occurrence of T. gondii infection could be detected. These findings suggest an environmental source of infection for mice associated with localized putative foci of environmental contamination and support an oral route of infection for mice from Dakar rather than a cycle based on vertical transmission. However, further investigations based on a denser sampling in different epidemiological contexts are recommended.

Prevalence and genetic characterization of Toxoplasma gondii in naturally infected backyard pigs intended for familial consumption in Romania.

BACKGROUND: Foodborne toxoplasmosis in humans can be due to the exposure to tissue cysts of Toxoplasma gondii through the consumption of meat, including pork, of infected animals. Traditional Romanian food habits include pork as the preferred meat, while backyard pig rearing remains a common practice in many rural areas of Romania. The aims of the present study were to estimate the prevalence of T. gondii infection in naturally infected backyard pigs slaughtered for familial consumption and to genetically characterize the T. gondii strains obtained. METHODS: Paired blood and heart samples were collected from 94 backyard pigs, home slaughtered for private consumption. Serum samples were analyzed using the immunofluorescence antibody test (IFAT) for anti-T. gondii antibody detection. Heart samples were screened by polymerase chain reaction (PCR) targeting the 529-bp repeat region (REP529) for T. gondii detection. In addition, heart samples from IFAT positive animals were bioassayed in mice. The T. gondii isolates were genotyped by the analysis of 15 microsatellite markers. RESULTS: The results showed that almost half of the pigs investigated were T. gondii seropositive (46.8%, 95% confidence interval (CI): 36.4-57.4%) and in more than a quarter of the pigs (26.6%, 95% CI: 18.0-36.7%), the parasite was detected by PCR. Three (3/44) T. gondii strains were isolated from hearts of seropositive pigs and they all belonged to genotype II. CONCLUSIONS: The present study showed the presence of T. gondii infection in backyard pigs in Romania, which suggests that consumption of pork from animals reared and slaughtered at home may pose a potential threat to human health and should be given attention. In addition, to our knowledge, this is the first study to provide data concerning T. gondii strains circulating in pigs from Romania.

The introduction of new hosts with human trade shapes the extant distribution of Toxoplasma gondii lineages.

Toxoplasma gondii is a zoonotic protozoan with a worldwide occurrence, but the determinants of the current pattern in the geographical distribution of T. gondii lineages and strains remain poorly understood. To test the influence of human trade on T. gondii populations, we conducted a population genetic study of 72 T. gondii animal isolates from Senegal, a West African country in which the ongoing inland progress of invasive murine hosts (introduced in port cities of Senegal since the 16th century by European sailors) is well described. Isolates were mainly collected on free-range poultry, which are considered as relevant bioindicators of T. gondii strain diversity in the domestic environment. Sampling was conducted in two port cities of Senegal (Dakar and Saint-Louis) and in one inland region (Kedougou). Population genetic analyses using 15 microsatellite markers revealed different patterns between port cities where lineages non-virulent for mice (type II, type III, and Africa 4) were predominant, and Kedougou where the mouse-virulent Africa 1 lineage was the most common. By considering the current spatial pattern in the inland progress of invasive rodents in Senegal, our results suggest that the invasive house mouse Mus musculus domesticus counter-selects the Africa 1 lineage in the invaded areas. The comparison of the microsatellite alleles of type II strains from Senegal to type II strains from other areas in Africa and Western Europe, using discriminant analysis of principal components and Network analysis, point to a mainly Western European origin of the type II lineage in Senegal. Collectively, these findings suggest that human-mediated intercontinental migrations of murine hosts are important vectors of T. gondii strains. Differential susceptibility of endemic and introduced murine hosts to various T. gondii strains probably determines the persistence of these strains in the environment, and therefore their availability for human and animal infection.

Exposure of yellow-legged gulls to Toxoplasma gondii along the Western Mediterranean coasts: Tales from a sentinel.

Efficiently tracking and anticipating the dynamics of infectious agents in wild populations requires the gathering of large numbers of samples, if possible at several locations and points in time, which can be a challenge for some species. Testing for the presence of specific maternal antibodies in egg yolks sampled on the colonies could represent an efficient way to quantify the exposure of breeding females to infectious agents, particularly when using an abundant and widespread species, such as the yellow-legged gull (Larus michahellis). We used such an approach to explore spatio-temporal patterns of exposure to Toxoplasma gondii, a pathogenic protozoan responsible of toxoplasmosis in humans and other warm-blooded vertebrates. First, we tested the validity of this approach by exploring the repeatability of the detection of specific antibodies at the egg level using two different immunoassays and at the clutch level using an occupancy model. Then, samples gathered in 15 colonies from France, Spain and Tunisia were analysed using an immunoassay detecting antibodies specifically directed against T. gondii. Prevalence of specific antibodies in eggs was overall high while varying significantly among colonies. These results revealed that T. gondii circulated at a large spatial scale in the western Mediterranean yellow-legged gull population, highlighting its potential role in the maintenance community of this parasite. Additionally, this study illustrates how species commensal to human populations like large gulls can be used as wildlife sentinels for the tracking of infectious agents at the human-wildlife interface, notably by sampling eggs.

Diversity of Toxoplasma gondii strains shaped by commensal communities of small mammals.

Commensal rodent species are key reservoirs for Toxoplasma gondii in the domestic environment. In rodents, different T. gondii strains show variable patterns of virulence according to host species. Toxoplasma gondii strains causing non-lethal chronic infections in local hosts will be more likely to persist in a given environment, but few studies have addressed the possible role of these interactions in shaping the T. gondii population structure. In addition, the absence of validated techniques for upstream detection of T. gondii chronic infection in wild rodents hinders exploration of this issue under natural conditions. In this study, we took advantage of an extensive survey of commensal small mammals in three coastal localities of Senegal, with a species assemblage constituted of both native African species and invasive species. We tested 828 individuals for T. gondii chronic infection using the modified agglutination test for antibody detection in serum samples and a quantitative PCR assay for detection of T. gondii DNA in brain samples. The infecting T. gondii strains were genotyped whenever possible by the analysis of 15 microsatellite markers. We found (i) a very poor concordance between molecular detection and serology in the invasive house mouse, (ii) significantly different levels of prevalence by species and (iii) the autochthonous T. gondii Africa 1 lineage strains, which are lethal for laboratory mice, only in the native African species of commensal small mammals. Overall, this study highlights the need to reconsider the use of MAT serology in natural populations of house mice and provides the first known data about T. gondii genetic diversity in invasive and native species of small mammals from Africa. In light of these results, we discuss the role of invasive and native species, with their variable adaptations to different T. gondii strains, in shaping the spatial structure of T. gondii genetic diversity in Africa.

Seroprevalence of Toxoplasma gondii in commensal rodents sampled across Senegal, West Africa.

Risks related to Toxoplasma gondii infection in humans remain poorly known in Senegal. Although rodent surveys could help to assess the circulation of T. gondii, they have seldom been set up in sub-Saharan Africa. The aim of this study was to examine Toxoplasma seroprevalence in rodents from villages and towns across Senegal. Rodents were sampled in 40 localities using a standardised trapping protocol. Detection of T. gondii antibodies was performed on 1205 rodents, using a modified agglutination test (MAT) technique. Seroprevalence data were analysed depending on geography, the local rodent community, and individual characteristics of the rodent hosts. We found 44 seropositive rodents from four different species (Mastomys erythroleucus, Mastomys natalensis, Mus musculus domesticus, Rattus rattus). Toxoplasma seroprevalence was low, averaging 4% in the localities. Higher Toxoplasma seroprevalence (up to 24%) was found in northern Senegal, a region known to be the heart of pastoral herding in the country.

TITLE: Séroprévalence de Toxoplasma gondii chez les rongeurs commensaux au Sénégal, Afrique de l'Ouest. ABSTRACT: Les risques liés à l'infection par Toxoplasma gondii chez l'homme restent mal connus au Sénégal. Bien que les rongeurs soient considérés comme des marqueurs pertinents de la circulation de T. gondii, peu d'études ont ciblé ces hôtes en Afrique sub-saharienne. Le but de cette étude était d'étudier la séroprévalence de Toxoplasma chez les rongeurs des villes et des villages du Sénégal. Les rongeurs ont été échantillonnés dans 40 localités avec un protocole de piégeage standardisé. La détection des anticorps contre T. gondii a été faite sur 1205 rongeurs avec un test d'agglutination modifié. Les données de séroprévalence ont été analysées en fonction de la géographie, de la communauté locale de rongeurs et de caractéristiques individuelles du rongeur. Nous avons trouvé 44 individus séropositifs, de quatre espèces différentes (Mastomys erythroleucus, Mastomys natalensis, Mus musculus domesticus, Rattus rattus). La séroprévalence moyenne est faible, de l'ordre de 4 % au sein des localités. Les plus fortes séroprévalences (jusqu'à 24 %) ont été observées dans le Nord du Sénégal, une région connue pour être le cœur de l'élevage pastoral dans le pays.