The impact of wildlife and environmental factors on hantavirus infection in the host and its translation into human risk.

Identifying factors that drive infection dynamics in reservoir host populations is essential in understanding human risk from wildlife-originated zoonoses. We studied zoonotic Puumala orthohantavirus (PUUV) in the host, the bank vole (Myodes glareolus), populations in relation to the host population, rodent and predator community and environment-related factors and whether these processes are translated into human infection incidence. We used 5-year rodent trapping and bank vole PUUV serology data collected from 30 sites located in 24 municipalities in Finland. We found that PUUV seroprevalence in the host was negatively associated with the abundance of red foxes, but this process did not translate into human disease incidence, which showed no association with PUUV seroprevalence. The abundance of weasels, the proportion of juvenile bank voles in the host populations and rodent species diversity were negatively associated with the abundance index of PUUV positive bank voles, which, in turn, showed a positive association with human disease incidence. Our results suggest certain predators, a high proportion of young bank vole individuals, and a diverse rodent community, may reduce PUUV risk for humans through their negative impacts on the abundance of infected bank voles.

Population- and Individual-Level Dynamics of the Intestinal Microbiota of a Small Primate.

UNLABELLED: Longitudinal sampling for intestinal microbiota in wild animals is difficult, leading to a lack of information on bacterial dynamics occurring in nature. We studied how the composition of microbiota communities changed temporally in free-ranging small primates, rufous mouse lemurs (Microcebus rufus). We marked and recaptured mouse lemurs during their mating season in Ranomafana National Park in southeastern mountainous rainforests of Madagascar for 2 years and determined the fecal microbiota compositions of these mouse lemurs with MiSeq sequencing. We collected 160 fecal samples from 71 animals and had two or more samples from 39 individuals. We found small, but statistically significant, effects of site and age on microbiota richness and diversity and effects of sex, year, and site on microbiota composition, while the within-year temporal trends were less clear. Within-host microbiota showed pervasive variation in intestinal bacterial community composition, especially during the second study year. We hypothesize that the biological properties of mouse lemurs, including their small body size and fast metabolism, may contribute to the temporal intraindividual-level variation, something that should be testable with more-extensive sampling regimes. IMPORTANCE: While microbiome research has blossomed in recent years, there is a lack of longitudinal studies on microbiome dynamics on free-ranging hosts. To fill this gap, we followed mouse lemurs, which are small heterothermic primates, for 2 years. Most studied animals have shown microbiota to be stable over the life span of host individuals, but some previous research also found ample within-host variation in microbiota composition. Our study used a larger sample size than previous studies and a study setting well suited to track within-host variation in free-ranging mammals. Despite the overall microbiota stability at the population level, the microbiota of individual mouse lemurs can show large-scale changes in composition in time periods as short as 2 days, suggesting caution in inferring individual-level patterns from population-level data.

Tracking year-to-year changes in intestinal nematode communities of rufous mouse lemurs (Microcebus rufus).

While it is known that intestinal parasite communities vary in their composition over time, there is a lack of studies addressing how variation in component communities (between-hosts) manifests in infracommunities (within-host) during the host lifespan. In this study, we investigate the changes in the intestinal parasite infracommunities in wild-living rufous mouse lemurs (Microcebus rufus) from Ranomafana National Park in southeastern Madagascar from 2010 to 2012. We used high-throughput barcoding of the 18S rRNA gene to interrogate parasite community structure. Our results show that in these nematode communities, there were two frequently occurring putative species and four rarer putative species. All putative species were randomly distributed over host individuals and they did not occur in clear temporal patterns. For the individuals caught in at least two different years, there was high turnover of putative species and high variation in fecal egg counts. Our study shows that while there was remarkable variation in infracommunities over time, the component community was relatively stable. Nevertheless, the patterns of prevalence varied substantially between years in each component community.

Relationships between learning strategies, stress, and study success among first-year veterinary students during an educational transition phase.

We investigated the relationships between stress, learning strategies, and study success among first-year veterinary students at the very beginning of their veterinary studies. The study was carried out during the first course on macroscopic anatomy (osteology), which students have in the past found to be exceptionally stressful. Students (N=45) completed a questionnaire concerning their self-reported views on stress and learning strategies, which were compared with their self-reported written-test scores. Participants who had previously gained university credits did not have significantly better test scores, but they achieved the learning goals with significantly less stress than other participants. Previous experience of university study helped students not only to adjust to a new type of course content and to achieve the learning goal of the osteology course, but also to cope with the stress experienced from taking concurrently running courses. Of the respondents who specifically named factors relating to self-regulation and modification of their learning strategy, all had gained prior credits. These students were able to use their study time efficiently and adjust their schedules according to the course demands.

Students as teachers in an anatomy dissection course.

One way to improve students' learning outcomes and well-being is to change teaching practices to allow students to become more active participants. We used an anatomy dissection course to test a cooperative group work method in which first-year veterinary students took turns leading their peer group and were each responsible for teaching the anatomy of a particular topographic region. The important blood vessels, lymphatic system, and nerves of each region of the body were covered. Students felt that exploration of the entire topographic region helped them to acquire a comprehensive understanding of the respiratory apparatus and the cardiovascular and nervous systems. Assigning individual tasks to each group member resulted in sharing the workload equally. Open-ended comments revealed that support from other group members was important for the students' learning experience, but the results also offered insight into a lack of constructive criticism. While teaching was considered challenging, and even a stress factor that hindered learning for some students, group work was generally held to be supportive of learning. The results suggest that more thorough instruction of students in their group work and in their individual tasks is required. Some students experienced difficulty in expressing their concerns openly and in seeking guidance from teachers, demonstrating the need for further investigation regarding students' self-regulation skills. Comments from the open-ended responses suggest that use of a cooperative learning method in anatomy dissection courses not only deepens student understanding of a subject but also offers first-year students an opportunity to practice the generic skills that will be needed in their future profession.

Comparison of chemical and enzymatic maceration processes for soft tissue removal from arterial silicone casts on skull scaffolds.

The various models used in gross anatomical studies to improve the visualization of blood vessels differ in the amount of manual labour, cost, equipment and time involved. This study aimed to compare chemical and enzymatic maceration processes for soft-tissue removal from arterial silicone casts on skull scaffolds using ringed seal (Pusa hispida) skull specimens. Both processes produced specimens that covered all anatomical aspects required to visualize the intracranial arterial arrangement on a bone scaffold. Overall, the enzyme maceration process was better for production of such specimens, as this process is easy and safe to perform, is less harmful to the bony parts of the specimen, and the resulting specimens are visually more appealing for display and teaching. Compared with previously published models, the end result varied in the amount of dissolved bone tissue and the visual presentation of the model.

Anatomical variations in the cerebral arterial circle of the Saimaa (Pusa hispida saimensis) and Baltic ringed seals (Pusa hispida botnica).

The intracranial arterial vascularization of the Saimaa ringed seals (Pusa hispida saimensis; Nordquist, 1899) and Baltic ringed seals (Pusa hispida botnica; Gmelin, 1788) disclosed patterns of anatomical architecture comparable to that of other pinniped species. Arterial silicone casts on skull scaffolds, and magnetic resonance imaging (MRI) showed that the besides joining the caudal communicating arteries upon entering the cerebral arterial circle, the bilateral internal carotid arteries bifurcated as laterally oriented rostral choroidal arteries and rostral cerebral arteries. The latter arteries almost immediately gave off the laterally oriented middle cerebral arteries. Numerous individual variations were evident in differences in the exact branching sites of bilateral vessels or the size or number of arterial branches. Two Saimaa ringed seals had only a tiny foramen for the left internal carotid artery to enter the intracranial space, and the intracranial part of this vessel was short. It did not reach the cerebral arterial circle. The intracranial part of the right internal carotid artery is bifurcated and also supplied the left side of the cerebral arterial circle. Both specimens had aplasia of the left rostral cerebral artery. The intracranial arterial arrangement of Saimaa and Baltic ringed seals reflects the arterial architecture of this body region in terrestrial mammals with little evidence for aquatic adaptations or changes related to thermoregulation.

Morphology of sweat glands in the skin of Saimaa (Pusa hispida saimensis) and Baltic ringed (Pusa hispida botnica) seals.

The endangered Saimaa ringed seal (Pusa hispida saimensis) is an endemic freshwater subspecies inhabiting Lake Saimaa in Finland. The Baltic ringed seal (Pusa hispida botnica) inhabits the brackish Baltic Sea, which is almost entirely landlocked. Recent research shows that Saimaa and Baltic ringed seals may be genetically even further apart from each other than from other ringed seal subspecies. We documented histologically the integument microstructure of Saimaa and Baltic ringed seals to determine whether the geographic and genetic isolation was manifested as variation in the integument microstructure of these subspecies adapted to icy aquatic environments. The skin structures of these subspecies were similar to those of other phocids. The association of the sweat glands with hair follicles in both subspecies suggested that they were small apocrine sweat glands described previously in terrestrial or aquatic mammals. None of the apocrine glands had large lumina, and some of the ducts were relatively straight and short. Further studies analysing the mode of secretion, for example, apocrine versus eccrine, in the sweat glands are necessary to confirm the types of sweat glands in seals.

Hip and stifle joint anatomy of the Saimaa (Pusa hispida saimensis) and Baltic ringed seals (Pusa hispida botnica).

The anatomical architecture of the hip and stifle joints of the Saimaa ringed seal was determined by dissection to be similar to that of the Baltic ringed seal. There was no ligament of the femoral head in the hip joint. The lateral side of the stifle joint lacked a collateral ligament but the collateral ligament on the medial side was of considerable size. The medial meniscus was larger and thicker than the lateral meniscus. The asymmetrical anatomical architecture allows for much lateral movement during aquatic motion, but it is poorly adapted to terrestrial locomotion.

Distribution and morphology of ventricular bands in the hearts of ringed seals.

In contrast to studies on domestic animals, few reports describe ventricular bands in wildlife, and none in aquatic mammals. Ventricular bands in the endangered Saimaa ringed seal (Pusa hispida saimensis) and the Baltic ringed seal (Pusa hispida botnica) were examined as part of an ongoing research on the comparative anatomy of ringed seal subspecies. The dissections illustrated that a varying number of thin or thick ventricular bands from the papillary muscles to the ventricular walls were visible in the ventricles of all ringed seal specimens examined. The histological appearance of the ventricular bands was characterized by a fibromuscular pattern.

Drawing in Veterinary Anatomy Education: What Do Students Use It For?

In contrast to medical education, information on the use of arts as a learning approach is scarce in veterinary anatomy. The aim of this prospective, questionnaire-based study was to survey students' use of drawing in various aspects of veterinary anatomy learning (self-study, examinations, preparation for, and reflection on cadaver dissection). The quantitative data showed that first-year students with artistic preferences used drawing significantly more often in most aspects of anatomy learning than students with no such preferences, apart from the reported use of drawing in examinations and for reflection after cadaver dissection. The lack of significant correlations of the reported use of drawing in examinations with any other study variable provided support for the author's observation that student-generated drawings are not as commonly used in examinations as previously. In contrast to the study hypothesis, previous university studies did not correlate significantly with any aspect of the use of drawing in anatomy learning. None of the reported uses of drawing addressed the benefits of drawing in learning the comparative anatomy of animal species, a characteristic distinguishing veterinary anatomy from human anatomy. Qualitative student feedback indicated that encouragement and teacher-produced images would increase the use of drawing as a learning approach even if the implementation of drawing into the curriculum is not feasible. Conclusions from this study were implemented through self-directed learning in anatomy courses that replaced the canceled cadaveric dissections during the COVID-19 outbreak and also through the ongoing drawing workshops to further advance the use of drawing in veterinary anatomy learning.

Unique hip and stifle extensor muscle patterns in the Eurasian lynx, Lynx lynx (Carnivora: Felidae).

The Eurasian lynx (Lynx lynx) is a medium-sized felid, with a tendency to hunt for prey larger than itself. We studied the lynx hindlimb musculoskeletal anatomy in order to determine possible anatomical adaptations to hunting large prey. In our previous work, we had found characters of both large and small felids in the lynx forelimb. The crouched limbs, typical of all felids, increase the energy demands for the antigravity muscles during locomotion. As a powerful pounce is required for the smaller felid to bring down large prey, strong hindquarters may be needed. We hypothesized that the muscle attachments are more mechanically advantageous and muscles heavier in the lynx as compared to other felids to compensate for the energy requirements. In support of this, we found unique patterns in the hindlimb musculature of the lynx. Insertion of the m. gluteus medius was large with a short moment arm around the hip joint, providing mechanical disadvantage, but rapid movement. The musculus vastus medialis was relatively heavier than in other felids emphasizing the role of the m. quadriceps femoris as a powerful stifle extensor. The extensor muscles support the crouched hind limbs, which is crucial when tackling large prey, and they are also responsible for the swift powerful pounce brought by extending the hindlimbs. However, we cannot rule out the possibility the characters are shared with other Lynx spp. or they are adaptations to other aspects of the locomotor strategy in the Eurasian lynx.

Estimating the amount of collagen and elastic fibres in bovine teats.

The amount of collagen and elastic fibres near the Fürstenberg's rosette in histological sections of bovine teats was estimated using the ImageJ image processing software. This method holds promise for comparing tissue types within and between sections but it was not a reliable way to quantify the absolute amount of tissue types in a sample. The amount of elastic fibres and collagen was similar in cow teats with a history or acute case of mastitis infection and in non-infected cows, but this could not be statistically tested due to limitations in the study material.

Spatial and temporal dynamics of lymphocytic choriomeningitis virus in wild rodents, northern Italy.

We determined the prevalence of infection with lymphocytic choriomeningitis virus (LCMV) among small mammals in northern Italy and analyzed long-term dynamics of LCMV in a rodent population in the province of Trento. LCMV is circulating among the most widespread and common wild rodent species in this area (Apodemus flavicollis, Myodes glareolus, and Microtus arvalis); overall prevalence is 6.8%. During 2000-2006, intensive monitoring of LCMV in a population of yellow-necked mice (A. flavicollis) showed a positive correlation between prevalence of infection and rodent density. At the individual level, weight and sex appeared to correlate with antibody prevalence, which suggests that horizontal transmission of LCMV occurs principally among heavier, older males and occurs during fighting. Isolation and genetic characterization of this virus will be the crucial next steps for a better understanding of its ecology.

Fostering Students' Collaborative Learning Competencies and Professional Conduct in the Context of Two Gross Anatomy Courses in Veterinary Medicine.

Collaborative learning has been used in anatomy courses to support students' learning of challenging topics but the success of group work depends significantly on the students' ability to communicate in a professional manner. Veterinary students' experiences with tasks related to collaborative learning and professional conduct were studied by comparing learning collaborative competences and pedagogy, as well as perceived positive and challenging aspects, in two gross anatomy courses. Both qualitative and quantitative data were collected from students' experiences of course assignments and collaboration, as well as from self-evaluated collaboration competence development. Trying things out oneself, practical application and professional conduct were positively highlighted particularly in the first-year course (myology and arthrology) group work and learning from others more in the second year (topographical anatomy). Various group work challenges, often relating to practical matters or communication, were the key concerns for students. The main difference between the two courses in learning of collaborative knowledge work competencies appeared to relate to the nature of the group work assignment. The topography course included the element of collaboration in preparing a presentation and teaching session for the rest of the class. Interestingly, students on the myology and arthrology course gave more positive comments on professional conduct than the students on the topography course despite the fact that the latter course included more practical elements relating to their future profession.

Association between the DQA MHC class II gene and Puumala virus infection in Myodes glareolus, the bank vole.

Puumala virus (PUUV) is a hantavirus specifically harboured by the bank vole, Myodes (earlier Clethrionomys) glareolus. It causes a mild form of hemorrhagic fever with renal syndrome (HFRS) in humans, called Nephropathia epidemica (NE). The clinical severity of NE is variable among patients and depends on their major histocompatibility complex (MHC) genetic background. In this study we investigated the potential role of class II MHC gene polymorphism in the susceptibility/resistance to PUUV in the wild reservoir M. glareolus. We performed an association study between the exon 2 of the DQA gene and PUUV antibodies considering a natural population of bank voles. Because immune gene polymorphism is likely to be driven by multiple parasites in the wild, we also screened bank voles for other potential viral and parasitic infections. We used multivariate analyses to explore DQA polymorphism/PUUV associations while considering the potential antagonist and/or synergistic effects of the whole parasite community. Our study suggests links between class II MHC characteristics and viral infections including PUUV and Cowpox virus. Several alleles are likely to be involved in the susceptibility or in the resistance of bank voles to these infections. Alternatively, heterozygosity does not seem to be associated with PUUV or any other parasite infections. This result thus provides no evidence in favour of the hypothesis of selection through overdominance. Finally this multivariate approach reveals a strong antagonism between ectoparasitic mites and PUUV, suggesting direct or indirect immunogenetic links between infections by these parasites. Other datasets are now required to confirm these results and to test whether the associations vary in space and/or time.

Genetic analysis of hantaviruses carried by Myodes and Microtus rodents in Buryatia.

Hantavirus genome sequences were recovered from tissue samples of Myodes rufocanus, Microtus fortis and Microtus oeconomus captured in the Baikal area of Buryatia, Russian Federation. Genetic analysis of S- and M-segment sequences of Buryatian hantavirus strains showed that Myodes-associated strains belong to Hokkaido virus (HOKV) type while Microtus-associated strains belong to Vladivostok virus (VLAV) type. On phylogenetic trees Buryatian HOKV strains were clustered together with M. rufocanus- originated strains from Japan, China and Far-East Russia (Primorsky region). Buryatian Microtus- originated strains shared a common recent ancestor with M. fortis- originated VLAV strain from Far-East Russia (Vladivostok area). Our data (i) confirm that M. rufocanus carries a hantavirus which is similar to but distinct from both Puumala virus carried by M. glareolus and Muju virus associated with M. regulus, (ii) confirm that M. fortis is the natural host for VLAV, and (iii) suggest M. oeconomus as an alternative host for VLAV.

Genetic analysis of Thailand hantavirus in Bandicota indica trapped in Thailand.

Sixty one tissue samples from several rodent species trapped in five provinces of Thailand were examined for the presence of hantaviral markers by enzyme-immunoassay and immunoblotting. Four samples, all from the great bandicoot rat Bandicota indica, were confirmed positive for the hantaviral N-antigen. Two of them were trapped in Nakhon Pathom province, the other two in Nakhon Ratchasima province, approximately 250 km from the other trapping site. When analysed by RT-nested PCR, all four rodents were found positive for the hantaviral S- and M-segment nucleotide sequences. Genetic analysis revealed that the four newly described wild-type strains belong to Thailand hantavirus. On the phylogenetic trees they formed a well-supported cluster within the group of Murinae-associated hantaviruses and shared a recent common ancestor with Seoul virus.

Effects of Protostrongylus sp. and Pneumocystis sp. on the pulmonary tissue and the condition of mountain and brown hares from Finland.

Mountain hares (Lepus timidus) and brown hares (Lepus europaeus) shot by hunters in several game management districts in southern and central Finland during the hunting season from September to the end of February 1998-2001 were examined for Protostrongylus sp. and Pneumocystis sp. Of the mountain hares, 96.5% (194/201) were infected with the lungworm Protostrongylus sp. and 16.9% (32/189) had cyst forms of the fungus Pneumocystis sp. in the lungs. The prevalence of the lungworm and fungus in brown hares was 60% (18/30) and 20.0% (6/30), respectively. The tissue changes associated with the lungworms were macroscopically and microscopically well demarcated. The majority and most severe histopathologic changes were seen at the distal part of the caudal lobes. Inflammatory cells, mainly eosinophils and macrophages, and in lesser degree neutrophils, lymphocytes, and plasma cells were typical findings in the worm-infected tissue. The condition and weight of the hare did not show any significant association with the intensity of the lungworm infection. All Pneumocystis-infected mountain hares were young, and their condition and weight correlated negatively with the intensity of the infection. The intensity of the Pneumocystis infection did not correlate with that of the lungworm infection. Within a tissue section, a slight but significant positive correlation was observed between presence of cysts and inflammatory cells.

Macroscopic Anatomy of the Saimaa Ringed Seal (Phoca hispida saimensis) Lower Respiratory Tract.

We studied the macroscopic anatomy of the lower respiratory tract of the endangered Saimaa ringed seal (Phoca hispida saimensis). Examination of one adult and one young individual found dead showed that trachea had 85 and 86 complete cartilage rings. The adjacent cartilages exhibited very few random anastomoses. There was variation in the confirmation of the trachea between the cranial and caudal part of the trachea. The right lung was divided by partly incomplete inter-lobar fissures into cranial, middle, caudal, and accessory lobes. The left lung consisted of cranial, middle, and caudal lobes. The lungs were characterized by a high amount of interlobular connective tissue. Silicone casts were prepared of the two specimens to visualize the tracheobronchial branching which was similar to that of marine ringed seals but in the Saimaa ringed seal the right middle lobar bronchus originated at the same level as the accessory lobar bronchus.