Optimized combination of Cervus nippon (Sika deer), Angelica (Dangui), and Rehmannia (Suk-jihwang) mitigates LPS-induced inflammation: exploring signaling pathways through plasma metabolomics.

This study aimed to determine the optimal combination of three anti-inflammatory materials [i.e., Cervus nippon Temminck (CT), Angelica gigas Nakai (AN), and Rehmannia glutinosa (RG)] for the strongest anti-inflammatory potential. Eighteen combinations of the three materials were tested in LPS-stimulated RAW264.7 cells via assessing nitric oxide (NO). The best combination from in vitro studies was administered to LPS-treated C57BL/6J mice for five days. Subsequently, plasma metabolites were profiled by bioinformatics analyses and validations. As results, 2, 20, and 50 µg/mL of CT, AN, and RG (TM) were the most effective combination suppressing inflammation. In mice, TM mitigated hepatic inflammatory markers. Similarly, the metabolomics indicated that TM may suppress NF-κB signaling pathway, thereby alleviating hepatic inflammation. TM also decreased systemic and hepatic pro-inflammatory cytokines. Collectively, we found the optimal combination of TM for mitigating inflammation; thus further studies on safety, mechanisms, and clinical models are warranted for human applications. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01476-x.

The role of large mammalian herbivores in shaping and maintaining soil microbial communities of natural mineral licks: A case study on sika deer at the firebreak adjacent to the Sino-Russian border.

Mineral licks are indispensable habitats to the life history of large mammal herbivores (LMH). Geophagy at licks may provide the necessary minerals for LMH, while LMH may be ecosystem engineers of licks by altering vegetation cover and soil physicochemical properties (SPCP). However, the precise relationship between the LMH and licks remains unclear. To clarify the geophagy function of licks for LMH and their influence on soil at licks, we recorded visitation patterns of sika deer around licks and compared SPCP and microbial communities with the surrounding matrix in a firebreak adjacent to the Sino-Russian border. Our study indirectly supports the "sodium supplementation" hypothesis. Proofs included (1) a significantly higher sodium, iron, and aluminum contents than the matrix, while lower carbon, nitrogen, and moisture contents; (2) significantly higher deer visitation during sodium-demand season (growing season), along with an avoidance of licks with high iron contents, which is toxic when overdose. The microbes at the licks differed from those at the matrix, mainly driven by low soil carbon and nitrogen and altered biogeochemical cycles. The microbial communities of licks are vulnerable because of their unstable state and susceptibility to SPCP changes. Structural equation modeling (SEM) clearly showed a much stronger indirect effect of deer on microbes at licks than at the matrix, especially for bacteria. Multiple deer behaviors at licks, such as grazing, trampling, and excretion, can indirectly shape and stabilize microbes by altering carbon and nitrogen input. Our study is the first to characterize soil microbial communities at mineral licks and demonstrate the processes by which LMH shapes those communities. More studies are required to establish a general relationship between the LMH and licks to promote the conservation of natural licks for wildlife.

Sleep with Open Eyes in Two Species of Deer, the Indian Sambar (Rusa unicolor) and Sika Deer (Cervus nippon).

The relationship between postures, sleep stages and eye state was established in two species of deer, the Indian sambar (Rusa unicolor) and sika deer (Cervus nippon), based on video recording. In both species, the state of rest or behavioral sleep was recorded in the sternal position, holding the head above the ground, and in the lateral position, with the head resting on the croup or on the ground. Rest accounted for at least 80% of the time in these positions. Based on behavior criteria a substantial portion of rest represented slow-wave sleep. Episodes of rapid eye movements (REM sleep) were recorded in the lateral position. They did not exceed 2 min. When the deer were in the sternal posture, they kept their eyes open most of the time: in average 96% of the time in sambars and 82% in sika deer. Episodes of the open eye in this posture lasted up to 8.4 min in sambars and up to 3.3 min in sika deer. In the lateral position, such episodes were 4 and 1.5 times shorter. Sleeping with open eyes in ungulates may be an important mechanism of maintaining vigilance.

Do Feed Plants Provide Sufficient Sodium, Calcium and Magnesium to Sika Deer in Japan? An Analysis Using Global Plant Trait Data.

Deficient minerals in overabundant populations could act as an attractant to cull sika deer (Cervus nippon). Because selective culling of female deer is reported to be effective in reducing sika deer populations, it is particularly important to clarify the differences in mineral requirements between males and females. Here, using global plant trait data and a published list of sika deer feed plants in Japan, we estimated whether feed plants provide sika deer sufficient sodium (Na), calcium (Ca) and magnesium (Mg), and compared the results between males and females. An analysis of 191 feed plant species suggested that feed plants can provide sufficient Mg, whereas sufficient Na and Ca is not always provided, especially when the intake is small or the deer large. Na deficiency was more intense for lactating females than males, suggesting that Na can be an effective attractant for selectively culling female deer. In summary, this study demonstrated that sika deer in Japan might require extra Na and Ca sources in addition to feed plants, and therefore these minerals could be useful for developing effective culling methods.

DNA methylation pattern and mRNA expression of OPN promoter in sika deer antler tip tissues.

In order to explore the biological role of OPN gene during the growth of sika deer antler, the dermis, mesenchyme, precartilage and cartilage tissues of sika deer antler tip at the early period of the antler with a saddle-like appearance (30 days), the rapid growth period of the antler with two branches (60 days), and the final period of the antler with three branches (90 days) were analyzed. Bisulfite sequencing PCR (BSP) and quantitative real-time PCR (qRT-PCR) were used to explore the DNA promoter methylation and mRNA expression of OPN in sika deer antler from the perspective of space and time. The test results showed that: 1) The methylation rates of OPN promoter at the early, middle and late periods of dermis tissue were (40.48 ± 0.82)%, (40.00 ± 1.43)%, and (39.05 ± 0.82)%; The methylation rates in mesenchyme tissue were (37.62 ± 0.82)%, (34.76 ± 2.18)%, and (38.57 ± 1.43)%; The methylation rates in precartilage tissue were (36.67 ± 0.28)%, (29.52 ± 1.65)%, (28.10 ± 2.18)%; The methylation rates in cartilage tissue were (31.90 ± 1.65)%, (26.67 ± 1.65)%, (24.29 ± 1.43)%. 2) There are 7 CpG sites in the OPN promoter region, and the 3 CpG sites of -367 bp, -245 bp and -31 bp are all methylated to different level. 3) The methylation level of OPN in the dermis, mesenchyme, precartilage and cartilage tissues decreased in sequence at the same growth period. At the middle and late periods, the methylation level of the promoter region of the precartilage tissue was significantly different from that of the dermis and mesenchyme tissues (P < 0.05); At different growth periods, the methylation level of the promoter region of cartilage tissue was extremely significantly different from that of dermis and mesenchyme tissues (P < 0.01); In the same tissue, the methylation level of the promoter region at the middle period was down-regulated compared with the early period, and the methylation level of the promoter region at the early period and the middle period was extremely significantly different in the precartilage and cartilage (P < 0.01). 4) OPN mRNA is highly expressed in precartilage and cartilage tissues. 5) The methylation level of OPN promoter was negatively correlated with mRNA expression level. In summary, it is speculated that the OPN gene, which may be regulated by the DNA methylation level of the promoter, promotes the growth and development of deer antler mainly by regulating the growth of precartilage and cartilage tissues.

Diet preferences based on sequence read count: The role of species interaction in tissue bias correction.

High-throughput sequencing and metabarcoding techniques provide a unique opportunity to study predator-prey relationships. However, in animal dietary preference studies, how to properly correct tissue bias within the sequence read count and the role of interactions between co-occurring species in metabarcoding mixtures remain largely unknown. In this study, we propose two categories of tissue bias correction indices: sequence read count number per unit tissue (SCN) and its ratio form (SCN ratio). By constructing plant mock communities with different numbers of co-occurring species in metabarcoding mixtures and conducting feeding trails on captive sika deer (Cervus nippon), we demonstrate the features of the SCN and SCN ratio, evaluate their correction effects and assess the role of species interactions during tissue bias correction. Tissue differences between species are defined as the differential ability to generate sequence counts. Our study suggests that pure tissue differences among species without a species interaction is not an optimal correction index for many biomes with limited tissue differences among species. Species interactions in mixtures may amplify tissue differences, which is beneficial for tissue bias correction. However, caution must be taken because varied species interactions among communities may increase the risk of worse correction. Correction effects based on the SCN and SCN ratio are comparable, but the SCN is less influenced by control species than the SCN ratio. Based on our study, several suggestions are provided for future animal diet studies or other high-throughput sequencing studies containing tissue bias.

Uncovering the Fecal Bacterial Communities of Sympatric Sika Deer (Cervus nippon) and Wapiti (Cervus canadensis).

Microbial symbiotic associations may be beneficial, neutral, or harmful to the host. Symbionts exploit the host space and nutrition or use hosts as carriers to spread to other environments. In order to investigate the fecal bacterial communities of wild sika deer (Cervus nippon) and wapiti (Cervus canadensis), this study aimed to sequence and explore the composition of, and similarity between, the fecal microbiota of sika deer and wapiti using high-throughput sequencing. The composition and relative abundance of fecal microbiota, alpha diversity, and differences in beta diversity between the two species were analyzed. We found that no pathogenic bacteria were present in large quantities in the hosts. The dominant bacterial phyla found in the two deer species were similar and included Firmicutes, Bacteroidetes, Proteobacteria, and Spirochaetes. Moreover, the deer also shared similar dominant genera, including the Rikenellaceae RC9 gut group, Ruminococcaceae_UCG-010, Ruminococcaceae_UCG-005, and Bacteroides. These results demonstrate that the sika deer and wapiti share a similar fecal microbiotal structure, probably due to their common diet and living environment, but there was some evidence of a difference at the species level. These analyses provide new insights into the health status of deer populations outside protected environments and offer a scientific framework for monitoring the health conditions of sika deer and wapiti.

Prevalence of Shiga toxin-producing Escherichia coli in Yezo sika deer (Cervus nippon yesoensis) in the Tokachi sub-prefecture of Hokkaido, Japan.

In food hygiene, the surveillance of foodborne pathogens in wild animals is indispensable because we cannot control hygienic status of them. Yezo sika deer (Cervus nippon yesoensis), which are found only on the island of Hokkaido, Japan, are the most common game animal in the country. In this study, we analyzed the incidence of Shiga toxin-producing Escherichia coli (STEC) in Yezo sika deer hunted in the Tokachi sub-prefecture, which is one of the densest zones for the sub-species. Real-time polymerase chain reaction testing detected STEC in 18.3% of fecal samples (59/323) collected from deer hunted between 2016 and 2017, whereas no Shigella and Salmonella markers were detected. No correlation was found between STEC detection from fecal samples and characteristics of carcasses, such as hunting area, age, and fascioliasis. From 59 STEC-positive fecal samples, we isolated 37 STEC strains, including 34 O- and H-genotyped strains, in which 16 different serogroups were detected. Genetic analysis revealed that our isolates included various stx gene types (stx1+/stx2-, stx1+/stx2+, and stx1-/stx2+) and carried eae. This study demonstrated that STEC strains with various features colonized the Yezo sika deer, similar to other subspecies of sika deer. We conclude that continuous surveillance activity is important to monitor the suitability of game animals as a food source and to assess the validity of the food safety management system for game meat production.

Genomic characterization and the prevalence of a novel copiparvovirus in wild sika deer (Cervus nippon) in Japan.

A preliminary metagenomic analysis of the virome of wild sika deer (Cervus nippon) blood in Japan resulted in the identification of a novel parvovirus. The virus was closest, but only 44.7-60.7% identical to 17 reported strains belonging to the genus Copiparvovirus within the subfamily Parvovirinae, over the near-entire genomic sequence. The sika deer copiparvovirus DNA was detected in 15% (31/206) of sika deer captured in 7 prefectures of Japan, and a region-dependent prevalence of 0-66.7% was noted, with a biased distribution in the southern part of Japan. The observed biased distribution of sika deer copiparvovirus may be due to the habitat density of deer and the number of ticks, which might play a role in the transmission of the virus.

Response of an Indicator Species, Dryopteris crassirhizoma, to Temporal and Spatial Variations in Sika Deer Density.

Identifying appropriate indicator species for the impact of deer on forest vegetation is crucial for forest management in deer habitats and is required to be sensitive to temporal and spatial variations in deer density. Dryopteris crassirhizoma was selected as a new indicator to evaluate the response to these variations. We examined the population-level characteristics, morphological characteristics at the individual level, and grazing intensity of D. crassirhizoma at temporally different deer density sites in Hokkaido, Japan. The response of D. crassirhizoma to spatial variation in deer density was also examined within and between two regions in Hokkaido, Japan. Although the population-level characteristics and morphological characteristics did not significantly respond to short-term decreases in deer density, grazing intensity significantly decreased with decreasing deer density. The grazing intensity was also positively related to the spatial variation of deer density within both regions, but the estimated coefficient of the grazing intensity differed between regions. We concluded that D. crassirhizoma can be a useful indicator species of the impact of deer on forest vegetation. The grazing intensity of the indicator species was sensitive to temporal and spatial variations in deer density within the region.

Sika Deer (Cervus nippon) are not "Dead-End Hosts" of the Giant Liver Fluke, Fascioloides magna (Bassi, 1875) Ward, 1917.

Fascioloides magna, although of North American origin, is well established in central Europe. Here it is expanding its geographic range and has been exposed to new potential final hosts including native and naturalized species. Based on their contribution to the propagation and transmission of F. magna, its final hosts have been grouped into three types including 'dead-end hosts' that comprise species in which flukes reach the liver but rarely mature and produce few eggs which are not excreted. Sika deer (Cervus nippon) were classified as dead-end hosts, this being quoted in the literature without robust substantiation. In an investigation on the occurrence of F. magna among sympatric wild ungulates in a recently established focus of the parasite in Germany, nine of 24 sika deer were found infected with up to seven F. magna in their livers, and six of the fluke-positive sika deer had Fascioloides eggs in their feces. Most flukes were recovered in pairs from fibrous capsules. Associated with the low fluke burden, gross pathology of the livers was generally mild. The presence of mature flukes in fibrous capsules, and passing of ova in the feces, demonstrates sika deer to be suitable definitive hosts of F. magna and to be of epidemiologic significance because of their implication in the transmission of the fluke.

Habitat Selection and Activity Patterns of Japanese Serows and Sika Deer with Currently Sympatric Distributions.

The Japanese serow (Capricornis crispus) and sika deer (Cervus nippon) in Japan are usually allopatric. However, a recent expansion in the distribution range of sika deer, combined with an increase in abundance, has resulted in an overlap of the distribution ranges of the two species. We examined the habitat selection and activity patterns of Japanese serows and sika deer using camera traps placed at 83 sites within a 210 km2 study area, where the distribution range of these two species has recently overlapped. Although both species were photographed throughout the study area, we observed a low spatial overlap between them. The Japanese serows selected steep slopes, and the sika deer selected areas far away from human settlements. In addition, the Japanese serows and sika deer exhibited temporal partitioning with diurnal and crepuscular activity patterns, respectively. The observed partitioning could be explained by differences in their species-specific habitat selections, rather than competition, because the photographic capture rate of the Japanese serows was not affected by that of the sika deer and vice versa. These partitioning behaviors are likely to reduce the rate of encounters between the two ungulates, which enables their coexistence considering the sika deer density remains low.

Occurrence patterns of crop-foraging sika deer distribution in an agriculture-forest landscape revealed by nitrogen stable isotopes.

Conflicts arising from the consumption of anthropogenic foods by wildlife are increasing worldwide. Conventional tools for evaluating the spatial distribution pattern of large terrestrial mammals that consume anthropogenic foods have various limitations, despite their importance in management to mitigate conflicts. In this study, we examined the spatial distribution pattern of crop-foraging sika deer by performing nitrogen stable isotope analyses of bone collagen. We evaluated whether crop-foraging deer lived closer to agricultural crop fields during the winter and spring, when crop production decreases. We found that female deer in proximity to agricultural crop fields during the winter and spring were more likely to be crop-foraging individuals. Furthermore, the likelihood of crop consumption by females decreased by half as the distance to agricultural crop fields increased to 5-10 km. We did not detect a significant trend in the spatial distribution of crop-foraging male deer. The findings of spatial distribution patterns of crop-foraging female deer will be useful for the establishment of management areas, such as zonation, for efficient removal of them.

Genetic diversity of cervid Trypanosoma theileri in Honshu sika deer (Cervus nippon) in Japan.

The taxonomy of ruminant Trypanosoma theileri and its relatives (Kinetoplastida: Trypanosomatidae) is controversial, with recent phylogenetic studies segregating T. theileri in cattle and other ruminants worldwide into two major genetic lineages (the TthI and TthII clades) based on genetic markers. In the present study, T. theileri-like trypanosomes isolated from Honshu sika deer (Cervus nippon) in the western Japan (YMG isolate) were genetically characterized using a number of genetic markers. Sika deer trypanosomes of the YMG isolate were genetically different from the Trypanosoma sp. TSD1 isolate previously recorded from Hokkaido sika deer in northern Japan, with the former trypanosome isolate being genetically closer to European cervid trypanosomes and the bovine T. theileri TthII lineage. In contrast, the latter isolate exhibited greater relatedness to North American cervid trypanosomes and the bovine T. theileri TthI lineage, although a clear genetic distinction between these was apparent. Furthermore, trypanosomes in Honshu sika deer from the central part of Japan harboured additional genetic diversity and were closer to either TSD1 or YMG isolates, while distinct from known T. theileri-related genotypes. Importantly, cervids and wild ruminants worldwide might harbour divergent descendants of a T. theileri ancestor, which exhibit rigid host specificity to either bovines or cervid species.

Protective effect of Sika Deer bone polypeptide extract on dexamethasone-induced osteoporosis in rats

BACKGROUND: Osteoporosis attacks approximately 10% of the population worldwide. Sika Deer (Cervus nippon), one of China's precious traditional medicinal animals, has been widely recorded in ancient Chinese medical books and claimed for centuries to have numerous medical benefits including bone strengthening. This study aimed to find the use of Sika Deer bone in treating osteoporosis according to traditional records and to investigate the protective effect of Sika Deer bone polypeptide extract on glucocorticoidinduced osteoporosis (GIOP) in rats. RESULTS: Sika Deer bone polypeptide extract could increase serum Ca2+ and BGP, decrease serum P3+, ALP, PTH, and CT, but had no effect on serum NO in rats with GIOP. The immunohistochemical iNOS results of the rats' distal femur were negative in each group. Besides the model group, the eNOS color reaction in osteoblasts was strongly positive in the other three groups. CONCLUSIONS: Sika Deer bone polypeptide extract can improve pathological changes in the microstructure and stimulate the expression of eNOS in osteoblasts. The protective effect on bone might be mediated by eNOS-dependent NO generation.

Bovine Tuberculosis: The Emergence of a New Wildlife Maintenance Host in Ireland.

Despite advances in herd management, tuberculosis (TB) continues to affect ~0. 5% of Ireland's national cattle herd annually. It is clear that any "final" eradication of TB in cattle will need to address all TB maintenance hosts in the same environment. In Ireland and the UK, European Badgers (Meles meles) are a known TB maintenance host, while deer are recognised as spillover hosts. However, deer have been identified as maintenance hosts in other countries and Sika deer, specifically, have been identified with TB in Ireland. We examined the power of cattle, badger and Sika deer densities (at the county level) to predict cattle TB-breakdowns in Ireland, at both the herd and the individual level, using data collected between 2000 and 2018. Our hypothesis was that any positive correlations between deer density and cattle TB-breakdowns would implicate deer as TB maintenance hosts. Using linear multiple regressions, we found positive correlations between deer density and cattle TB-breakdowns at both the herd and individual levels. Since Sika deer in County Wicklow are known to have TB, we ran further regressions against subsets of data which excluded individual Irish counties. Analyses excluding Wicklow data showed much weaker correlations between Sika deer density and cattle TB-breakdowns at both the herd and individual levels, suggesting that these correlations are strongest in County Wicklow. A similar effect for badger density was seen in County Leitrim. While locally high densities of Sika deer persist in Irish counties, we believe they should be considered an integral part of any TB-control programme for those areas.

Species-specific responses of a marsh-forest ecotone plant community responding to climate change.

Ecotones are responsive to environmental change and pave a path for succession as they move across the landscape. We investigated the biotic and abiotic filters to species establishment on opposite ends of a tidal marsh-forest ecotone that is moving inland in response to sea level rise. We transplanted four plant species common to the ecotone to the leading or trailing edge of the migrating ecotone, with and without caging to protect them from ungulate herbivores. We found that species exhibited an individualistic response to abiotic and biotic pressures in this ecotone; three species performed better at the leading edge of the ecotone in the coastal forest, whereas one performed better at the trailing edge in the marsh. Specifically, grass species Phragmites australis and Panicum virgatum grew more in the low light and low salinity conditions of the leading edge of the ecotone (forest), whereas the shrub Iva frutescens grew better in the high light, high salinity conditions of the trailing edge of the ecotone (marsh). Furthermore, of the four species, only P. australis was affected by the biotic pressure of herbivory by an introduced ungulate, Cervus nippon, which greatly reduced its biomass and survival at the leading edge (forest). P. australis is an aggressive invasive species and has been observed to dominate in the wake of migrating marsh-forest ecotones. Our findings detail the role of lower salinity stress to promote and herbivory pressure to inhibit the establishment of P. australis during shifts of this ecotone, and also highlight an interaction between two nonnative species, P. australis and C. nippon. Understanding migration of the marsh-forest ecotone and the factors controlling P. australis establishment are critical for marsh conservation in the face of sea level rise. More generally, our findings support the conclusion that the abiotic and biotic filters of a migrating ecotone shape the resulting community.

Clonal Spread of Quinolone-Resistant Escherichia coli among Sika Deer (Cervus nippon) Inhabiting an Urban City Park in Japan.

The emergence and dissemination of resistance to clinically important antimicrobials in wild animals is of great concern. The aim of our study was to reveal the prevalence and intraspecies dissemination of quinolone-resistant Escherichia coli (QREC) in sika deer (Cervus nippon) in Nara Park, a famous tourist spot in Japan. Fecal samples were collected from 59 wild deer in Nara Park between July and October 2018. We isolated QREC using deoxycholate-hydrogen sulfide-lactose agar containing nalidixic acid and subjected it to antimicrobial susceptibility testing. The mutations in the quinolone resistance-determining region (QRDR) of the gyrA and parC genes of the isolates were analyzed and fragment patterns of genomic DNA were compared by pulsed-field gel electrophoresis (PFGE). A total of 105 QREC isolates were obtained from 41 deer (70%). All isolates had mutations within the QRDR. Other than quinolone resistance, QREC isolates also showed resistance to various other antimicrobial agents. The QREC isolates were classified into 15 PFGE clusters, of which seven were observed in multiple deer. Our results suggest clonal transmission of QREC in a high-density deer population. Spread of QREC in deer inhabiting a tourist location could have potential impact on public health.

Phylogeography of sika deer (Cervus nippon) inferred from mitochondrial cytochrome-b gene and microsatellite DNA.

The genetic diversity and phylogenetic relationships of sika deer of different subspecies are uncertain. In order to explore the phylogenetic relationship of different sika deer subspecies, this study used a wider sample collection to analyze mitochondrial sequences and nuclear microsatellites of sika deer. The full lengths of cytochrome-b gene of 134 sika deer were sequenced, and 16 haplotypes were obtained. Based on phylogenetic and haplotype networks analysis, the sika deer was not clustered according to subspecies but was divided into four lineages. Lineage I includes individuals from C.n.kopschi, C.n.sichuanicus, and C.n.hortulorum subspecies; Lineage II includes individuals from C.n.hortulorum subspecies; Lineage III includes individuals from C.n.centralis, C.n.yakushime, C.n.mageshimae, and C.n.keramae subspecies, namely southern Japanese population; Lineage IV includes individuals from C.n.centralis and C.n.yesoensis subspecies, namely northern Japanese population. The microsatellite analysis showed that the sika deer in China and Japan originated independently. The three subspecies of China have significant genetic differentiation, while the three subspecies of Japan have no significant differentiation. This study provides reference for the research of genetic diversity and phylogenetic relationship of sika deer, and also provides scientific data for the evaluation, protection, and utilization of sika deer resources.

Anaplasma phagocytophilum and Babesia Species of Sympatric Roe Deer (Capreolus capreolus), Fallow Deer (Dama dama), Sika Deer (Cervus nippon) and Red Deer (Cervus elaphus) in Germany.

(1) Background: Wild cervids play an important role in transmission cycles of tick-borne pathogens; however, investigations of tick-borne pathogens in sika deer in Germany are lacking. (2) Methods: Spleen tissue of 74 sympatric wild cervids (30 roe deer, 7 fallow deer, 22 sika deer, 15 red deer) and of 27 red deer from a farm from southeastern Germany were analyzed by molecular methods for the presence of Anaplasma phagocytophilum and Babesia species. (3) Results: Anaplasma phagocytophilum and Babesia DNA was demonstrated in 90.5% and 47.3% of the 74 combined wild cervids and 14.8% and 18.5% of the farmed deer, respectively. Twelve 16S rRNA variants of A. phagocytophilum were delineated. While the infection rate for A. phagocytophilum among the four cervid species was similar (71.4% to 100%), it varied significantly for Babesia between roe deer (73.3%), fallow deer (14.3%), sika deer (27.3%) and red deer (40.0%). Deer ≤2 years of age tested significantly more often positive than the older deer for both A. phagocytophilum and Babesia species. (4) Conclusions: This study confirms the widespread occurrence of A. phagocytophilum and Babesia species in wild cervids and farmed red deer in Germany and documents the co-occurrence of the two tick-borne pathogens in free-ranging sika deer.