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As part of this project, we here focus on a serologic survey of rodent-borne Toxoplasma , which concerns a sub-total of 46 trapping sites Figure , Supplementary data. However, for the purpose Supplementary data as well as Figure , they were aggregated for a clearer visualisation at the whole town scale. Rodents were live-trapped and brought to the lab where they were usually euthanised within one-eight days, except for 45 of them which were autopsied within eight days data not shown.
All procedures were carried out in agreement to current ethical guidelines for animal care. The blotting papers were dried and stored in a plastic bag at room temperature RT. All details are provided in Garba Serological survey of T. Two 5 mm diameter dried blood spot were punched out of each blotting paper circle and placed into the well of a flat bottomed microtitre plate. Plates were covered and left to elute overnight at RT and at rpm agitation.
Ten microlitres of each eluted sample was used in MAT technique. For serological control, fresh blood from seronegative not infected by T. Antibody titres were determined by the last dilution where agglutination pattern could be read in comparison with the negative and positive controls.
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Statistical analysis - Descriptive analyses of the serological data were based on frequencies, percentage for qualitative variables and means, standard deviations for quantitative variables. Relationships between rodent seroprevalence and factors such as sex, species and habitats were investigated through chi-squared test or Fisher's exact test, depending on the expected sample size.
For each significant factor, a Cochran-Mantel-Haenszel Chi-Squared test was conducted in order to obtain a p-value adjusted for the other factors. All statistics were performed using the software R v2. From the 46 trapping sites sampled for the present study, rodents could be surveyed for Toxoplasma serology: were black rats Rattus rattus , 61 were house mice M.
Two animals possessed atypical profiles not shown and so were fully sequenced for their cytochrome b mitochondrial gene. These DNA sequences allowed to barcode them and to unambiguously identify them as M. These unambiguously identified M. Moreover, they originated from all 30 trapping sites were Mastomys individuals were trapped Supplementary data. We can therefore conclude that all Mastomys that were trapped in the present survey belong to one single species, namely M. Seropositive individuals represent 1. They belong to four different species: the native M. No seropositive individuals were found in C.
Seropositive individuals included both juvenile and adult animals as well as males and females Table III. In most instances, they corresponded to one single seropositive specimen found within five specimens from one particular trapping site; only in two exceptions CGA and J-LMO did we find several seropositive animals within the same trapping site Table III , Supplementary data.
Finally, in three instances, seropositive individuals were part of multiple captures i. Represented rodent species are typical Sahelian species that were all already known in the area Dobigny et al. Although differentiating between rural and urban environments in Niamey may sometimes be tricky since the two types of habitats are often continuous when not fully intermingled houses closely surrounding or lying within gardens and rice fields, gardens within familial concessions etc , rodent species distribution in regards to biotopes was quite clear: A. Global T. This parallels previous studies where positive urban rodents are usually rare.
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For instance, surveys in Brazilian cities showed 4. As for other studies, the seroprevalence results may be discussed according to the sensitivity and specificity of the serological test. Seroprevalence in our study was evaluated through a modified-agglutination test which is the most commonly used for defining a possible infection in diverse species of animals, as there is no need for specific secondary antibodies.
The most commonly considered cut-off is , but T. That explains why we choose the cut-off of that represents the lowest dilution available after elution of dried blood spot. The gold-standard for detection of T. This was not possible in the context of Niger. PCR-based method for Toxoplasma DNA detection on tissue samples brain, muscles is known to have a lower sensitivity than bioassay and serology Hill et al.
This once again fits to what was observed for rodents elsewhere in the World, with species-specific seroprevalence ranging from close to null e. It is also noteworthy that the same rodent species can display extremely different seroprevalence depending on localities or countries. For instance, seroprevalence in R. These specific as well as geographic variations point toward a complex T. Seropositive rodents were recorded across the year Table III , encompassing all of the Sahelian seasons [from the warm and dry season March and April , through the rainy season June , to the cool and dry season October and November ], thus suggesting that Toxoplasma infection may occur throughout the year in Niamey's rodents.
However, diachronic monitoring within the same site was not feasible, thus precluding any conclusion about potential seasonal seroprevalence peaks. Although our data are limited both in nature we score antibodies, not proper infection cases and sample size, we can rely on three instances of multiple hence simultaneous captures to partly address this point Table III.
Unfortunately, no data about any adult is available here, thus making it impossible to decipher between independent environmental infections - for example at the same place, such as the nest - and vertical transmission. Third, a triple capture included a seronegative adult female and two seropositive juveniles of A.
These two latter cases bring poor support though not refute to vertical transmission and rather suggest that animals get infected from the environment soil and water. Interestingly, the more typical commensal species found in Niamey M. In particular, only six individuals of the native and widespread species in Niamey, i. This species is found within houses in all investigated parts of the city. These cats mainly survive from garbage and wild preys, something that may maximize the risk for them to get infected by ingestion of infected rodents.
Low seroprevalence in Mastomys which is, from far, the dominating species in most habitations, hence the most susceptible to be a major cats' prey may limit cat predation-mediated infections through commensal rodents, hence in turn decreasing potential transmissions from cats to humans. Another important aspect for public health is the similarly low seroprevalences observed in M.
Indeed, these two invasive species recently established in Niamey Garba and it is possible that their populations may potentially extend within the city, potentially partly replacing the native M. Bovine and ovine meat is traditionally well cooked in Niger. In addition, rodent meat consumption by humans is rather rare in Niger, especially in Niamey and most exclusively concerns young boys that occasionally hunt in gardens.
Also, previous studies conducted in Nigeria Olusi et al. For all these reasons, following previous authors Develoux et al. Low levels of T. This most probably also limits the chance of environmental contaminations.
Nevertheless, such extreme and unfavourable conditions may be locally counteracted by human-mediated modifications of the habitat. In particular, the possibly major role of direct waterborne contamination has been receiving increasing support [e. In the absence of other feasible explanations, water was even speculated as a major source of toxoplasmic infection in pregnant women and children from Northern Niger Dumas et al.
Interestingly, we found here significantly higher seroprevalence in A. Unfortunately, we were not able to statistically address this particular issue here, since we could not decipher between Arvicanthis -specific epidemiological properties and environmental i. Indeed, food habits are clearly switching towards higher consumption of vegetables e. Rodents such as Nile grass rats feed mainly on the cultivated vegetables Dobigny et al. It was previously shown that risk of infection in French cats was higher with warm and moist weather Afonso et al.
In Niamey, temperatures are always high the coldest month January is characterised by a normal minimal temperature of As such, the connection in Sahelian cities between oocysts, watered vegetables and rodents could be a key element of Toxoplasma circulation that may deserve to be further scrutinised. To our knowledge, the present study is the first one to focus on T. In spite of a large sample size, seroprevalence was found to be rather low, with a possible exception in A.
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