Perinatal mortality in German dairy cattle: Unveiling the importance of cow-level risk factors and their interactions using a multifaceted modelling approach.

Perinatal mortality (PM) is a common issue on dairy farms, leading to calf losses and increased farming costs. The current knowledge about PM in dairy cattle is, however, limited and previous studies lack comparability. The topic has also primarily been studied in Holstein-Friesian cows and closely related breeds, while other dairy breeds have been largely ignored. Different data collection techniques, definitions of PM, studied variables and statistical approaches further limit the comparability and interpretation of previous studies. This article aims to investigate the factors contributing to PM in two underexplored breeds, Simmental (SIM) and Brown Swiss (BS), while comparing them to German Holstein on German farms, and to employ various modelling techniques to enhance comparability to other studies, and to determine if different statistical methods yield consistent results. A total of 133,942 calving records from 131,657 cows on 721 German farms were analyzed. Amongst these, the proportion of PM (defined as stillbirth or death up to 48 hours of age) was 6.1%. Univariable and multivariable mixed-effects logistic regressions, random forest and multimodel inference via brute-force model selection approaches were used to evaluate risk factors on the individual animal level. Although the balanced random forest did not incorporate the random effect, it yielded results similar to those of the mixed-effect model. The brute-force approach surpassed the widely adopted backwards variable selection method and represented a combination of strengths: it accounted for the random effect similar to mixed-effects regression and generated a variable importance plot similar to random forest. The difficulty of calving, breed and parity of the cow were found to be the most important factors, followed by farm size and season. Additionally, four significant interactions amongst predictors were identified: breed-calving ease, breed-season, parity-season and calving ease-farm size. The combination of factors, such as secondiparous SIM breed on small farms and experiencing easy calving in summer, showed the lowest probability of PM. Conversely, primiparous GH cows on large farms with difficult calving in winter exhibited the highest probability of PM. In order to reduce PM, appropriate management of dystocia, optimal heifer management and a wider use of SIM in dairy production are possible ways forward. It is also important that future studies are conducted to identify farm-specific contributors to higher PM on large farms.

Benchmarking calf health: Assessment tools for dairy herd health consultancy based on reference values from 730 German dairies with respect to seasonal, farm type, and herd size effects.

Good calf health is crucial for a successfully operating farm business and animal welfare on dairy farms. To evaluate calf health on farms and to identify potential problem areas, benchmarking tools can be used by farmers, herd managers, veterinarians, and other advisory persons in the field. However, for calves, benchmarking tools are not yet widely established in practice. This study provides hands-on application for on-farm benchmarking of calf health. Reference values were generated from a large dataset of the "PraeRi" study, including 730 dairy farms with a total of 13,658 examined preweaned dairy calves. At herd level, omphalitis (O, median 15.9%) was the most common disorder, followed by diarrhea (D, 15.4%) and respiratory disease (RD, 2.9%). Abnormal weight bearing (AWB) was rarely detected (median, 0.0%). Calves with symptoms of more than one disorder at the same time (multimorbidity, M) were observed with a prevalence of 2.3%. The enrolled farms varied in herd size, farm operating systems, and management practices and thus represented a wide diversity in dairy farming, enabling a comparison with similar managed farms in Germany and beyond. To ensure comparability of the data in practice, the reference values were calculated for the whole data set, clustered according to farm size (1-40 dairy cows (n = 130), 41-60 dairy cows (n = 99), 61-120 dairy cows (n = 180), 121-240 dairy cows (n = 119) and farms with more than 240 dairy cows (n = 138), farm operating systems (conventional (n = 666), organic (n = 64)) and month of the year of the farm visit. There was a slight tendency for smaller farms to have a lower prevalence of disorders. A statistically significant herd-size effect was detected for RD (p = 0.008) and D (p < 0.001). For practical application of these reference values, tables, diagrams, and an Excel® (Microsoft®) based calf health calculator were developed as tools for on-farm benchmarking (https://doi.org/10.6084/m9.figshare.c.6172753). In addition, this study provides a detailed description of the colostrum, feeding and housing management of preweaned calves in German dairy farms of different herd sizes and farm type (e.g., conventional and organic).

Retrospective evaluation of vector-borne pathogens in cats living in Germany (2012-2020).

BACKGROUND: Blood-feeding arthropods can transmit parasitic, bacterial, or viral pathogens to domestic animals and wildlife. Vector-borne infections are gaining significance because of increasing travel and import of pets from abroad as well as the changing climate in Europe. The main objective of this study was to assess the percentage of cats with positive test results for selected vector-borne pathogens in Germany and explore any possible association of such results with time spent abroad. METHODS: This retrospective study included test results from cats included in the "Feline Travel Profile" established by the LABOKLIN laboratory at the request of veterinarians in Germany between April 2012 and March 2020. This diagnostic panel includes the direct detection of Hepatozoon spp. and Dirofilaria spp. via PCR as well as indirect detection assays (IFAT) for Ehrlichia spp. and Leishmania spp. The panel was expanded to include an IFAT for Rickettsia spp. from July 2015 onwards. RESULTS: A total of 624 cats were tested using the "Feline Travel Profile." Serum for indirect detection assays was available for all 624 cats; EDTA samples for direct detection methods were available from 618 cats. Positive test results were as follows: Ehrlichia spp. IFAT 73 out of 624 (12%), Leishmania spp. IFAT 22 out of 624 (4%), Hepatozoon spp. PCR 53 out of 618 (9%), Dirofilaria spp. PCR 1 out of 618 cats (0.2%), and Rickettsia spp. IFAT 52 out of 467 cats (11%) tested from July 2015 onwards. Three cats had positive test results for more than one pathogen before 2015. After testing for Rickettsia spp. was included in 2015, 19 cats had positive test results for more than one pathogen (Rickettsia spp. were involved in 14 out of these 19 cats). CONCLUSIONS: At least one pathogen could be detected in 175 out of 624 cats (28%) via indirect and/or direct detection methods. Four percent had positive test results for more than one pathogen. These data emphasize the importance of considering the above-mentioned vector-borne infections as potential differential diagnoses in clinically symptomatic cats.

Retrospective evaluation of vector-borne infections in dogs imported from the Mediterranean region and southeastern Europe (2007-2015).

BACKGROUND: Canine vector-borne infections have gained importance in Germany due to growing tourist traffic and an increased import of dogs from abroad. Endemic regions for pathogens such as Leishmania infantum, Hepatozoon canis, Ehrlichia canis, Anaplasma platys and Dirofilaria spp. are the Mediterranean area and southeastern Europe. Babesia species and Anaplasma phagocytophilum are present all over Europe. The objective of this retrospective study was to evaluate the prevalence of vector-borne infections in dogs imported from defined endemic countries in the Mediterranean area and southeastern Europe. METHODS: Medical records and laboratory test results of 345 dogs that were imported to Germany from 17 endemic countries and that were presented to the Small Animal Clinic at Freie Universität Berlin between 2007 and 2015 were retrospectively reviewed. A total of 1368 test results from external laboratories were descriptively analysed including 576 and 792 test results of direct and indirect detection methods, respectively. RESULTS: Overall, 35% (122/345 dogs) were positive for at least one pathogen. Concurrent infections with two to four pathogens were detected in 8% of the dogs (27/345). The positive results were: L. infantum 21% (66/314 dogs; methods: PCR 20/79, IFAT or ELISA 63/308 dogs), E. canis 16% (45/278 dogs; methods: PCR 8/68, IFAT 43/257 dogs), H. canis 11% (3/28 dogs; method: PCR), Babesia spp. 10% (25/251 dogs; methods: Babesia spp. PCR 3/98, B. canis/vogeli IFAT or ELISA 22/214 and B. gibsoni IFAT 0/13 dogs), Dirofilaria spp. 7% (13/178 dogs; methods: D. immitis Ag-ELISA 8/156, Knott's test 7/95, microfilariae PCR 5/23 dogs) and A. platys 5% (1/21 dogs; method: PCR). None of 8 tested dogs were positive in a combined Babesia spp./Hepatozoon spp. PCR test. CONCLUSIONS: Dogs, which are imported from countries which are endemic for vector-borne infections should be thoroughly tested using direct and indirect detection methods. Potential owners of imported dogs should be informed about the diseases, risks and incubation periods.

Retrospective analysis of vector-borne infections in dogs after travelling to endemic areas (2007-2018).

Canine vector-borne infections gained in importance in Germany due to growing tourist traffic, the increased import of dogs from abroad and the changing of climatic conditions. The Mediterranean region and southeastern Europe are geographical areas where pathogens such as Leishmania (L.) infantum, Hepatozoon (H.) canis, Ehrlichia (E.) canis, Anaplasma (A.) platys and Dirofilaria (D.) spp. are endemic. Meanwhile, Babesia (B.) spp. and A. phagocytophilum are present in central and western Europe. The objective of this retrospective study was to evaluate whether dogs were exposed to a corresponding risk of infection when travelling to regions in the Mediterranean area and southeastern Europe, which are endemic for these pathogens. Medical records and laboratory test results of 303 dogs that travelled to 14 countries endemic for the mentioned canine vector-borne pathogens and that were presented to the Small Animal Clinic at Freie Universität Berlin between 2007 and 2018 were retrospectively reviewed. A total of 1174 test results from external laboratories were descriptively analysed including 525 test results of direct and 649 of indirect determination methods. Overall, 13% of the tested dogs (40/303) were positive for at least one pathogen. Concurrent infections with two pathogens were detected in 1% of the dogs (4/303). The positive results were: E. canis 8% (18/231 dogs; Polymerase chain reaction [PCR] 3/73, indirect immunofluorescence test [IFAT] 18/209 dogs), L. infantum 5% (14/260 dogs; PCR 5/80, IFAT or enzyme linked immunosorbent assay [ELISA] 11/251 dogs), Babesia spp. 5% (11/232 dogs; Babesia spp. PCR 3/127, B. canis/vogeli IFAT or ELISA 8/160, B. gibsoni IFAT 2/22), Dirofilaria spp. 1% (1/133 dogs; D. immitis antigen-ELISA 1/117, microfilariae PCR 0/16, Knott´s test 0/69 dogs). None of the dogs has been tested positive in a combined Babesia spp./Hepatozoon spp. PCR test (0/15 dogs) and for H. canis (0/17 dogs; PCR) or A. platys (0/11 dogs; PCR). There is a substantial risk for dogs travelling to areas endemic for vector-borne pathogens even with limited time of exposure to get infected. The data indicates the importance of owner education and prophylactic measurements against vector-borne infections in dogs travelling to endemic areas.

Retrospective analysis of vector-borne infections in dogs after travelling to endemic areas (2007-2018).

Canine vector-borne infections gained in importance in Germany due to growing tourist traffic, the increased import of dogs from abroad and the changing of climatic conditions. The Mediterranean region and southeastern Europe are geographical areas where pathogens such as Leishmania (L.) infantum, Hepatozoon (H.) canis, Ehrlichia (E.) canis, Anaplasma (A.) platys and Dirofilaria (D.) spp. are endemic. Meanwhile, Babesia (B.) spp. and A. phagocytophilum are present in central and western Europe. The objective of this retrospective study was to evaluate whether dogs were exposed to a corresponding risk of infection when travelling to regions in the Mediterranean area and southeastern Europe, which are endemic for these pathogens. Medical records and laboratory test results of 303 dogs that travelled to 14 countries endemic for the mentioned canine vector-borne pathogens and that were presented to the Small Animal Clinic at Freie Universität Berlin between 2007 and 2018 were retrospectively reviewed. A total of 1174 test results from external laboratories were descriptively analysed including 525 test results of direct and 649 of indirect determination methods. Overall, 13% of the tested dogs (40/303) were positive for at least one pathogen. Concurrent infections with two pathogens were detected in 1% of the dogs (4/303). The positive results were: E. canis 8% (18/231 dogs; Polymerase chain reaction [PCR] 3/73, indirect immunofluorescence test [IFAT] 18/209 dogs), L. infantum 5% (14/260 dogs; PCR 5/80, IFAT or enzyme linked immunosorbent assay [ELISA] 11/251 dogs), Babesia spp. 5% (11/232 dogs; Babesia spp. PCR 3/127, B. canis/vogeli IFAT or ELISA 8/160, B. gibsoni IFAT 2/22), Dirofilaria spp. 1% (1/133 dogs; D. immitis antigen-ELISA 1/117, microfilariae PCR 0/16, Knott´s test 0/69 dogs). None of the dogs has been tested positive in a combined Babesia spp./Hepatozoon spp. PCR test (0/15 dogs) and for H. canis (0/17 dogs; PCR) or A. platys (0/11 dogs; PCR). There is a substantial risk for dogs travelling to areas endemic for vector-borne pathogens even with limited time of exposure to get infected. The data indicates the importance of owner education and prophylactic measurements against vector-borne infections in dogs travelling to endemic areas.

Fecal S100A12 concentration predicts a lack of response to treatment in dogs affected with chronic enteropathy.

S100A12 is a potential biomarker of gastrointestinal inflammation in dogs and fecal S100A12 concentrations are correlated with disease severity and outcome. The aim of the present study was to investigate whether there was any association between pre-treatment fecal S100A12 concentrations in dogs affected with chronic enteropathy (CE) and the response to treatment. Dogs affected with CE were recruited into the study and were classified as antibiotic-responsive diarrhea (ARD; n = 9), food-responsive diarrhea (FRD; n = 30) or idiopathic inflammatory bowel disease (IBD; n = 25). They were also grouped based on their response to treatment as complete remission (n = 35), partial response (n = 25) or no response (n = 4). Fecal S100A12 concentrations, measured by ELISA, were elevated in dogs affected with IBD compared with those from dogs affected with FRD (P = 0.010) or ARD (P = 0.025). Dogs with IBD that did not respond to treatment (n = 4) had significantly greater fecal S100A12 concentrations than dogs in complete remission (P = 0.009). Measurement of fecal S100A12 at the time of diagnosis discriminated between dogs with IBD that were refractory to therapy (≥2700 ng/g fecal S100A12) from those with at least a partial response (<2700 ng/g fecal S100A12), with a sensitivity of 100% and a specificity of 76%. These preliminary results suggest that testing of fecal S100A12 may be useful for predicting the lack of response to treatment in dogs affected with CE. The utility of serial fecal S100A12 measurements for monitoring dogs undergoing treatment for CE warrants further investigation.

Frequencies of regulatory T cells in the peripheral blood of dogs with primary immune-mediated thrombocytopenia and chronic enteropathy: a pilot study.

Regulatory T (Treg) cells are specialized immune cells with a pivotal role in the maintenance of immune homeostasis and control of inflammation. However, relatively little is known about immune regulation in the peripheral blood of dogs with primary immune-mediated thrombocytopenia or chronic enteropathy. Using flow cytometry this study investigated Treg responses in the peripheral blood of dogs with respective autoimmune or chronic intestinal diseases and demonstrated that a reduction of Treg frequencies is observed in dogs with clinical signs compared to dogs undergoing remission and healthy dogs. These findings suggest that reduced frequencies of Treg cells in peripheral blood might be causally associated with the onset and/or progression of autoimmune and chronic intestinal diseases in dogs and that measurement of regulatory T cells might represent a useful tool in the monitoring of treatment response and disease progression.

Orbus hercynius gen. nov., sp. nov., isolated from faeces of wild boar, is most closely related to members of the orders 'Enterobacteriales' and Pasteurellales.

A novel gammaproteobacterium, strain CN3(T), was isolated from the faeces of wild boar. Strain CN3(T) was facultatively anaerobic and appeared coccoid or rod-shaped. The partial 16S rRNA gene sequence determined for strain CN3(T) suggested a distant relationship with members of the orders 'Enterobacteriales' and Pasteurellales. The gene sequence showed highest similarity (90.3 %) with Obesumbacterium proteus DSM 2777(T), a member of the family Enterobacteriaceae. The closest relatives outside the order 'Enterobacteriales' according to 16S rRNA gene sequence analysis were members of the order Pasteurellales with 88.7 % similarity (Mannheimia haemolytica NCTC 9380(T) and Actinobacillus lignieresii NCTC 4189(T)). In contrast to most members of the order 'Enterobacteriales', strain CN3(T) was oxidase-positive. The pattern of fatty acids, in particular the high relative abundance of C(18 : 1)ω7c (38.5 %), was clearly distinct from the conserved pattern found for members of the order Pasteurellales. EcoRI ribotyping of strain CN3(T) yielded no significant similarity to existing database entries. The major ubiquinone of strain CN3(T) was Q-8. The DNA G+C content was 36.4 mol%. Strain CN3(T) hosted a phage and secreted considerable amounts of three proteins into the culture supernatant. A spontaneous mutant of strain CN3(T) was isolated which formed long filaments. Microscopic studies revealed the presence of a capsule that the mutant strain was unable to partition after cell division. Strain CN3(T) thus represents a novel species within a new genus, for which the name Orbus hercynius gen. nov., sp. nov. is proposed. The type strain of the type species is CN3(T) (=DSM 22228(T)=CCUG 57622(T)). Classification of the novel species to the family and order level will require further investigations.