The abundance and distribution of sandflies (with emphasis on Phlebotomus pedifer) (Diptera: Psychodidae) along the altitudinal gradient in Kindo Didaye district, Wolaita Zone, South Ethiopia.

Understanding the distribution patterns of medically significant sandflies is crucial for effective vector and disease control planning. This study focused on investigating the abundance and distribution of phlebotomine sandflies, specifically emphasizing Phlebotomus pedifer (Diptera: Psychodidae), the vector of Leishmania aethiopica responsible for cutaneous leishmaniasis in the highlands of southern Ethiopia. The study employed CDC light traps and sticky paper traps in various habitats, including human houses, farm fields, and rock cliffs, with and without the presence of hyraxes. The study was conducted along an altitudinal gradient in Kindo Didaye district, Wolaita Zone. A total of 7,994 sandflies belonging to 2 genera, Phlebotomus (26.1%) and Sergentomyia (73.9%), were collected. In the genus Phlebotomus, P. pedifer (74.1%) was the most abundant, followed by P. alexandri (18.05%) and P. gibiensis (7.85%). Altitude showed a strong positive association with the density and distribution of P. pedifer and a negative association with those of P. alexandri, P. gibiensis, and Sergentomyia spp. Furthermore, the study revealed distinct habitat preferences, with P. pedifer showing the highest mean density in hyrax dwellings, followed by human houses, and the lowest in farm fields. These findings provide valuable insights for planning targeted control measures against P. pedifer in both indoor and outdoor environments, particularly in the highland and midland areas of the study region.

Twenty-nine years of continuous monthly capture-mark-recapture data of multimammate mice (Mastomys natalensis) in Morogoro, Tanzania.

The multimammate mice (Mastomys natalensis) is the most-studied rodent species in sub-Saharan Africa, where it is an important pest species in agriculture and carrier of zoonotic diseases (e.g. Lassa virus). Here, we provide a unique dataset that consists of twenty-nine years of continuous monthly capture-mark-recapture entries on one 3 ha mosaic field (MOSA) in Morogoro, Tanzania. It is one of the most accurate and long-running capture-recapture time series on a small mammal species worldwide and unique to Africa. The database can be used by ecologists to test hypotheses on the population dynamics of small mammals (e.g. to test the effect of climate change), or to validate new algorithms on real long-term field data (e.g. new survival analyses techniques). It is also useful for both scientists and decision-makers who want to optimize rodent control strategies and predict outbreaks of multimammate mice.

SARS-CoV-2 Surveillance between 2020 and 2021 of All Mammalian Species in Two Flemish Zoos (Antwerp Zoo and Planckendael Zoo).

The COVID-19 pandemic has led to millions of human infections and deaths worldwide. Several other mammal species are also susceptible to SARS-CoV-2, and multiple instances of transmission from humans to pets, farmed mink, wildlife and zoo animals have been recorded. We conducted a systematic surveillance of SARS-CoV-2 in all mammal species in two zoos in Belgium between September and December 2020 and July 2021, in four sessions, and a targeted surveillance of selected mammal enclosures following SARS-CoV-2 infection in hippopotamuses in December 2021. A total of 1523 faecal samples from 103 mammal species were tested for SARS-CoV-2 via real-time PCR. None of the samples tested positive for SARS-CoV-2. Additional surrogate virus neutralisation tests conducted on 50 routinely collected serum samples from 26 mammal species were all negative. This study is the first to our knowledge to conduct active SARS-CoV-2 surveillance for several months in all mammal species of a zoo. We conclude that at the time of our investigation, none of the screened animals were excreting SARS-CoV-2.

Drivers behind co-occurrence patterns between pathogenic bacteria, protozoa, and helminths in populations of the multimammate mouse, Mastomys natalensis.

Advances in experimental and theoretical work increasingly suggest that parasite interactions within a single host can affect the spread and severity of wildlife diseases. Yet empirical data to support predicted co-infection patterns are limited due to the practical challenges of gathering convincing data from animal populations and the stochastic nature of parasite transmission. Here, we investigated co-infection patterns between micro- (bacteria and protozoa) and macroparasites (gastro-intestinal helminths) in natural populations of the multimammate mouse (Mastomys natalensis). Fieldwork was performed in Morogoro (Tanzania), where we trapped 211 M. natalensis and tested their behaviour using a modified open-field arena. All animals were checked for the presence of helminths in their gastro-intestinal tract, three bacteria (Anaplasma, Bartonella, and Borrelia) and two protozoan genera (Babesia and Hepatozoon). Besides the presence of eight different helminth genera (reported earlier), we found that 19% of M. natalensis were positive for Anaplasma, 10% for Bartonella, and 2% for Hepatozoon species. Hierarchical modelling of species communities was used to investigate the effect of the different host-related factors on these parasites' infection probability and community structure. Our results show that the infection probability of Bartonella increased with the host's age, while the infection probability of Anaplasma peaked when individuals reached adulthood. We also observed that less explorative and stress-sensitive individuals had a higher infection probability with Bartonella. Finally, we found limited support for within-host interactions between micro-and macroparasites, as most co-infection patterns could be attributed to host exposure time.

Knowledge, attitude, and practice of the rural community about cutaneous leishmaniasis in Wolaita zone, southern Ethiopia.

BACKGROUND: Cutaneous leishmaniasis (CL) is a neglected tropical disease that is caused by a Leishmania parasite and transmitted by the bite of infected female sandflies. Community awareness is an essential component of disease control and prevention. Therefore, this study aimed to assess the community's knowledge, attitude, and practice toward CL in Wolaita zone, southern Ethiopia. METHODS: A community-based cross-sectional study design was employed to include 422 study subjects selected using a systematic sampling technique from two districts, Kindo Didaye and Sodo Zuria. A pretested structured questionnaire was used to collect data from the household heads. Bivariate and multivariate logistic regression analyses were performed to determine the relationship between the participants' knowledge about CL and socio-demographic characteristics. RESULTS: Out of the 422 study participants, only 19% had good knowledge of CL in general. Most (67.1%) of the respondents knew CL by its local name ("bolbo" or "moora") though this knowledge varied highly over the study districts. The majority (86.3%) of respondents did not know how CL is acquired, though they considered CL a health problem. Most (62.8%) respondents believed that CL was an untreatable disease. Most (77%) participants responded that CL patients preferred to go to traditional healers for treatment. Herbal treatment was the most (50.2%) used to treat CL. Knowledge about CL was significantly associated with sex, age, and study districts. CONCLUSION: The overall knowledge, attitude, and practice about CL and its prevention in the study area were low. This emphasizes the need to implement health education and awareness campaign to reduce the risk of CL infection. Policymakers and stakeholders should also give due attention to the prevention and treatment of CL in the study area.

Subspecific rodent taxa as the relevant host taxonomic level for mammarenavirus host specificity.

Mastomys natalensis-borne mammarenaviruses appear specific to subspecific M. natalensis taxa rather than to the whole species. Yet mammarenaviruses carried by M. natalensis are known to spill over and jump hosts in northern sub-Saharan Africa. Phylogeographic studies increasingly show that, like M. natalensis, small mammals in sub-Saharan Africa are often genetically structured into several subspecific taxa. Other mammarenaviruses may thus also form virus-subspecific host taxon associations. To investigate this, and if mammarenaviruses carried by M. natalensis in southern Africa are less prone to spill-over, we screened 1225 non-M. natalensis samples from Tanzania where many small mammal taxa meet. We found mammarenavirus RNA in 6 samples. Genetic/genomic characterisation confirmed they were not spill-over from M. natalensis. We detected host jumps among rodent tribe members and an association between mammarenaviruses and subspecific taxa of Mus minutoides and Grammomys surdaster, indicating host genetic structure may be crucial to understand virus distribution and host specificity.

SARS-CoV-2 Infection in Captive Hippos (Hippopotamus amphibius), Belgium.

Two adult female hippos in Zoo Antwerp who were naturally infected with SARS-CoV-2 showed nasal discharge for a few days. Virus was detected by immunocytochemistry and PCR in nasal swab samples and by PCR in faeces and pool water. Serology was also positive. No treatment was necessary.

Assessing animal welfare impact of fourteen control and dispatch methods for house mouse (Mus musculus), Norway rat (Rattus norvegicus) and black rat (Rattus rattus).

Population control of the house mouse (Mus musculus), Norway rat (Rattus norvegicus) and black rat (Rattus rattus) is common practice worldwide. Our objective was to assess the impact on animal welfare of lethal and non-lethal control methods, including three dispatch methods. We used the Sharp and Saunders welfare assessment model with eight experts scoring eleven control methods and three dispatch methods used on the three species. We presumed the methods were performed as prescribed, only taking into account the effect on the target animal (and not, for example, on non-target catches). We did not assess population control efficacy of the methods. Methods considered to induce the least suffering to the target animal were captive-bolt traps, electrocution traps and cervical dislocation, while those with the greatest impact were anticoagulants, cholecalciferol and deprivation. Experts indicated considerable uncertainty regarding their evaluation of certain methods, which emphasises the need for further scientific research. In particular, the impact of hydrogen cyanide, chloralose and aluminium phosphide on animal welfare ought to be investigated. The experts also stressed the need to improve Standard Operating Procedures and to incorporate animal welfare assessments in Integrated Pest Management (IPM). The results of our study can help laypeople, professionals, regulatory agencies and legislators making well-informed decisions as to which methods to use when controlling commensal rodents.

Quantitative assessment of the dispersal of soil-dwelling oribatid mites via rodents in restored heathlands.

Heathland restoration using topsoil removal requires the re-colonization of above- and belowground communities. Oribatid mites play a key role in the comminution of organic matter and are frequently early colonizers during succession despite their limited mobility. Whereas the assembly of their communities may take decades, passive dispersal likely dominates colonization processes, but especially dispersal via other animals (phoresy) remains poorly studied. Compared to other potential hosts, movement habits and ecology of small rodents may provide dispersal advantages to oribatid communities. We studied dispersal of oribatid mites via small rodents in restored heathland sites of different age. We measured movement patterns of small rodents and extracted mites from their pelts and nests to estimate annual contributions of these rodents to the dispersal of oribatids. We also discussed phoretic estimates reported on other host groups as a reference. Probability estimates of oribatids in pelts and nests showed lower occurrence frequencies compared to other reported phoretic hosts. However, local rodent communities may aid the dispersal of up to 41,000 oribatid mites per year. We highlight the high diversity of oribatid species mounting rodents, unlike strong species-specific filters reported in other passive pathways. We found that over half (58%) of the oribatid species reproduced asexually and over a third (32%) had a soil-dwelling lifestyle. We also observed that rodents often travel short distances below 40 m, but occasionally reach distances of up to 100 m, especially in earlier successional stages. Synthesis and applications. Our results suggest that rodents may contribute to assembly processes of soil-dwelling oribatid communities given the slow turnover rate of this group in heathlands. This is accomplished through short-distance dispersal, and especially in sites at early stages of succession. To our knowledge, we are the first to quantitatively assess the potential dispersal of oribatid mites via rodents.

Serologic Surveillance for SARS-CoV-2 Infection among Wild Rodents, Europe.

We report results from serologic surveillance for exposure to SARS-CoV-2 among 1,237 wild rodents and small mammals across Europe. All samples were negative, with the possible exception of 1. Despite suspected potential for human-to-rodent spillover, no evidence of widespread SARS-CoV-2 circulation in rodent populations has been reported to date.Esitämme tulokset serologisesta tutkimuksesta, jossa seulottiin SARS-CoV-2 tartuntojen varalta 1,237 luonnonvaraista jyrsijää ja piennisäkästä eri puolilta Eurooppaa. Kaikki näytteet olivat negatiivisia, yhtä näytettä lukuun ottamatta. SARS-CoV-2:n läikkymisen ihmisistä jyrsijöihin on arveltu olevan mahdollista, mutta todisteet viruksen laajamittaisesta leviämisestä jyrsijäpopulaatioissa puuttuvat.

Enhanced surveillance of monkeypox in Bas-Uélé, Democratic Republic of Congo: the limitations of symptom-based case definitions.

BACKGROUND: Following an outbreak of cases of vesicular-pustular rash with fever, evocative of human monkeypox, in Bas-Uélé province, Democratic Republic of Congo, surveillance was strengthened. METHODS: Households with at least one active generalized vesicular-pustular rash case were visited, and contact and clinical history information were collected from all household members. Whenever possible, skin lesions were screened by polymerase chain reaction for the monkeypox virus, followed by the varicella-zoster virus, when negative for the former. RESULTS: Polymerase chain reaction results were obtained for 77 suspected cases, distributed in 138 households, of which 27.3% were positive for monkeypox, 58.4% positive for chickenpox, and 14.3% negative for both. Confirmed monkeypox cases presented more often with monomorphic skin lesions on the palms of the hands and on the soles of the feet. Integrating these three features into the case definition raised the specificity to 85% but would miss 50% of true monkeypox cases. A predictive model fit on patient demographics and symptoms had 97% specificity and 80% sensitivity but only 80% and 33% in predicting out-of-sample cases. CONCLUSION: Few discriminating features were identified and the performance of clinical case definitions was suboptimal. Rapid field diagnostics are needed to optimize worldwide early detection and surveillance of monkeypox.

SARS-CoV-2 surveillance in Norway rats (Rattus norvegicus) from Antwerp sewer system, Belgium.

SARS-CoV-2 human-to-animal transmission can lead to the establishment of novel reservoirs and the evolution of new variants with the potential to start new outbreaks in humans. We tested Norway rats inhabiting the sewer system of Antwerp, Belgium, for the presence of SARS-CoV-2 following a local COVID-19 epidemic peak. In addition, we discuss the use and interpretation of SARS-CoV-2 serological tests on non-human samples. Between November and December 2020, Norway rat oral swabs, faeces and tissues from the sewer system of Antwerp were collected to be tested by RT-qPCR for the presence of SARS-CoV-2. Serum samples were screened for the presence of anti-SARS-CoV-2 IgG antibodies using a Luminex microsphere immunoassay (MIA). Samples considered positive were then checked for neutralizing antibodies using a conventional viral neutralization test (cVNT). The serum of 35 rats was tested by MIA showing three potentially positive sera that were later negative by cVNT. All tissue samples of 39 rats analysed tested negative for SARS-CoV-2 RNA. This is the first study that evaluates SARS-CoV-2 infection in urban rats. We can conclude that the sample of rats analysed had never been infected with SARS-CoV-2. However, monitoring activities should continue due to the emergence of new variants prone to infect Muridae rodents.

Linking Behavior, Co-infection Patterns, and Viral Infection Risk With the Whole Gastrointestinal Helminth Community Structure in Mastomys natalensis.

Infection probability, load, and community structure of helminths varies strongly between and within animal populations. This can be ascribed to environmental stochasticity or due to individual characteristics of the host such as their age or sex. Other, but understudied, factors are the hosts' behavior and co-infection patterns. In this study, we used the multimammate mouse (Mastomys natalensis) as a model system to investigate how the hosts' sex, age, exploration behavior, and viral infection history affects their infection risk, parasitic load, and community structure of gastrointestinal helminths. We hypothesized that the hosts' exploration behavior would play a key role in the risk for infection by different gastrointestinal helminths, whereby highly explorative individuals would have a higher infection risk leading to a wider diversity of helminths and a larger load compared to less explorative individuals. Fieldwork was performed in Morogoro, Tanzania, where we trapped a total of 214 individual mice. Their exploratory behavior was characterized using a hole-board test after which we collected the helminths inside their gastrointestinal tract. During our study, we found helminths belonging to eight different genera: Hymenolepis sp., Protospirura muricola, Syphacia sp., Trichuris mastomysi, Gongylonema sp., Pterygodermatites sp., Raillietina sp., and Inermicapsifer sp. and one family: Trichostrongylidae. Hierarchical modeling of species communities (HMSC) was used to investigate the effect of the different host-related factors on the infection probability, parasite load, and community structure of these helminths. Our results show that species richness was higher in adults and in females compared to juveniles and males, respectively. Contrary to our expectations, we found that less explorative individuals had higher infection probability with different helminths resulting in a higher diversity, which could be due to a higher exposure rate to these helminths and/or behavioral modification due to the infection.

Molecular detection and genomic characterization of diverse hepaciviruses in African rodents.

Hepatitis C virus (HCV; genus Hepacivirus) represents a major public health problem, infecting about three per cent of the human population. Because no animal reservoir carrying closely related hepaciviruses has been identified, the zoonotic origins of HCV still remain unresolved. Motivated by recent findings of divergent hepaciviruses in rodents and a plausible African origin of HCV genotypes, we have screened a large collection of small mammals samples from seven sub-Saharan African countries. Out of 4,303 samples screened, eighty were found positive for the presence of hepaciviruses in twenty-nine different host species. We, here, report fifty-six novel genomes that considerably increase the diversity of three divergent rodent hepacivirus lineages. Furthermore, we provide strong evidence for hepacivirus co-infections in rodents, which were exclusively found in four sampled species of brush-furred mice. We also detect evidence of recombination within specific host lineages. Our study expands the available hepacivirus genomic data and contributes insights into the relatively deep evolutionary history of these pathogens in rodents. Overall, our results emphasize the importance of rodents as a potential hepacivirus reservoir and as models for investigating HCV infection dynamics.

Prevalence of Orthohantavirus-Reactive Antibodies in Humans and Peri-Domestic Rodents in Northern Ethiopia.

In 2012, Tigray orthohantavirus was discovered in Ethiopia, but its seasonal infection in small mammals, and whether it poses a risk to humans was unknown. The occurrence of small mammals, rodents and shrews, in human inhabitations in northern Ethiopia is affected by season and presence of stone bunds. We sampled small mammals in two seasons from low- and high-density stone bund fields adjacent to houses and community-protected semi-natural habitats in Atsbi and Hagere Selam, where Tigray orthohantavirus was first discovered. We collected blood samples from both small mammals and residents using filter paper. The presence of orthohantavirus-reactive antibodies in blood was then analyzed using immunofluorescence assay (human samples) and enzyme linked immunosorbent assays (small mammal samples) with Puumala orthohantavirus as antigen. Viral RNA was detected by RT-PCR using small mammal blood samples. Total orthohantavirus prevalence (antibodies or virus RNA) in the small mammals was 3.37%. The positive animals were three Stenocephalemys albipes rats (prevalence in this species = 13.04%). The low prevalence made it impossible to determine whether season and stone bunds were associated with orthohantavirus prevalence in the small mammals. In humans, we report the first detection of orthohantavirus-reactive IgG antibodies in Ethiopia (seroprevalence = 5.26%). S. albipes lives in close proximity to humans, likely increasing the risk of zoonotic transmission.

Detection of Cutaneous Leishmaniasis Foci in South Ethiopia.

Cutaneous leishmaniasis (CL) is a major public health problem in Ethiopia. The disease is endemic in Ochollo, a village in southern Ethiopia, but there are no reports of CL in the wider area, although it is ecologically very similar. We conducted a rapid assessment survey in the South Ethiopian Rift Valley and found 100 parasitologically confirmed CL cases in 38 villages not reported endemic for CL. Approximately half of the cases were children (57%), and most lesions occurred on the face (78%) and were older than 6 months (77%). Only 2% of the people was aware of the mode of transmission, and 9% sought modern treatment at a hospital. These preliminary data indicate that CL is much more widespread than previously reported and that the disease might have a large psychosocial impact. Hence, this study calls for larger surveys across the Ethiopian highlands. Additionally, health education and treatment capacity need to be implemented.

Effects of forest disturbance on the fitness of an endemic rodent in a biodiversity hotspot.

Praomys delectorum occurs abundantly in both disturbed and intact forests in the Ukaguru Mountains within the Eastern Arc Mountains (EAM), Morogoro, Tanzania. While previous studies have reported that anthropogenic disturbances such as grazing, wood cutting, and harvesting have a positive effect on the population density of P. delectorum, the impact of habitat disturbance on its demographic traits is still unknown. We performed a capture-mark-recapture study in both disturbed and intact forests from June 2018 to February 2020 in order to investigate the effects of habitat disturbance on abundance and two demographic traits: survival and maturation of P. delectorum in the Ukaguru Mountains. We found no variation in abundance or maturation between intact and disturbed forests, but habitat type did affect survival. However, this effect was sex-dependent since female survival was higher in disturbed forests, while male survival remained similar across the two forest types potentially due to differences in predation pressure or food availability between the two habitats. Continuous demographic monitoring of P. delectorum in EAM is necessary given that the increasing human population surrounding the landscape is leading to higher deforestation rates and expansion of the pine plantation in the forest reserve.

Nonlinear maternal effects on personality in a rodent species with fluctuating densities.

Consistent among-individual variation in behavior, or animal personality, is present in a wide variety of species. This behavioral variation is maintained by both genetic and environmental factors. Parental effects are a special case of environmental variation and are expected to evolve in populations experiencing large fluctuations in their environment. They represent a non-genetic pathway by which parents can transmit information to their offspring, by modulating their personality. While it is expected that parental effects contribute to the observed personality variation, this has rarely been studied in wild populations. We used the multimammate mouse Mastomys natalensis as a model system to investigate the potential effects of maternal personality on offspring behavior. We did this by repeatedly recording the behavior of individually housed juveniles which were born and raised in the lab from wild caught females. A linear correlation, between mother and offspring in behavior, would be expected when the personality is only affected by additive genetic variation, while a more complex relationship would suggests the presence of maternal effects. We found that the personality of the mother predicted the behavior of their offspring in a non-linear pattern. Exploration behavior of mother and offspring was positively correlated, but only for slow and average exploring mothers, while this correlation became negative for fast exploring mothers. This may suggests that early maternal effects could affect personality in juvenile M. natalensis, potentially due to density-dependent and negative frequency-dependent mechanisms, and therefore contribute to the maintenance of personality variation.

The effects of personality on survival and trappability in a wild mouse during a population cycle.

The pace-of-life syndrome (POLS) theory provides an evolutionary explanation for the existence of consistent among-individual variation in behaviour, or animal personality. Herein, individuals with a fast lifestyle are considered to be bolder and should take more risks resulting in a lower life expectancy compared to shyer individuals with a slower lifestyle. However, this assumption depends on the levels of intra-specific competition that the individuals experience which has rarely been tested in species that experience large changes in competition on a very short time scale. We used the multimammate mice (Mastomys natalensis) as a model system to study the POLS assumption by investigating the effects of two personality traits (exploration and stress-sensitivity) on survival, maturation (a proxy for reproductive investment) and recapture probability during one population cycle (Nindividuals = 201). Such a cycle consists of two phases in which the levels of intra-specific competition vary drastically. We found that only one personality trait, namely stress-sensitivity, had a negative effect on both survival and recapture probability but none of them affected maturation. This suggests that less stress-sensitive individuals take more risks in the wild and have a higher survival probability compared to high stress-sensitive individuals. However, the effect of personality on survival was only present during the population decrease phase, when the levels of intra-specific competition are high due to a scarcity of food. This suggests that seasonal changes in competition might be important in the evolution and maintenance of animal personalities in species whose population dynamics have a clear seasonal component.

Evaluation of conventional and four real-time PCR methods for the detection of Leishmania on field-collected samples in Ethiopia.

In most low-resource settings, microscopy still is the standard method for diagnosis of cutaneous leishmaniasis, despite its limited sensitivity. In Ethiopia, the more sensitive molecular methods are not yet routinely used. This study compared five PCR methods with microscopy on two sample types collected from patients with a suspected lesion to advise on optimal diagnosis of Leishmania aethiopica. Between May and July 2018, skin scrapings (SS) and blood exudate from the lesion spotted on filter paper (dry blood spot, DBS) were collected for PCR from 111 patients of four zones in Southern Ethiopia. DNA and RNA were simultaneously extracted from both sample types. DNA was evaluated by a conventional PCR targeting ITS-1 and three probe-based real-time PCRs: one targeting the SSU 18S rRNA and two targeting the kDNA minicircle sequence (the 'Mary kDNA PCR' and a newly designed 'LC kDNA PCR' for improved L. aethiopica detection). RNAs were tested with a SYBR Green-based RT-PCR targeting spliced leader (SL) RNA. Giemsa-stained SS smears were examined by microscopy. Of the 111 SS, 100 were positive with at least two methods. Sensitivity of microscopy, ITS PCR, SSU PCR, Mary kDNA PCR, LC kDNA PCR and SL RNA PCR were respectively 52%, 22%, 64%, 99%, 100% and 94%. Microscopy-based parasite load correlated well with real-time PCR Ct-values. Despite suboptimal sample storage for RNA detection, the SL RNA PCR resulted in congruent results with low Ct-values. DBS collected from the same lesion showed lower PCR positivity rates compared to SS. The kDNA PCRs showed excellent performance for diagnosis of L. aethiopica on SS. Lower-cost SL RNA detection can be a complementary high-throughput tool. DBS can be used for PCR in case microscopy is negative, the SS sample can be sent to the referral health facility where kDNA PCR method is available.