Establishment of a Single Temperature Incubation Approach for Environmental Monitoring Samples with Focus on Mold Recoveries.

A robust environmental monitoring program is essential to properly estimate and identify microorganisms in cleanrooms, ensuring that microbial contamination remains acceptably low, and a good state of control is maintained in the manufacturing areas. The incubation conditions are important to support optimal microbial recoveries, considering that there is no single culture medium, temperature and incubation time that can recover all microorganisms. Particularly molds are quite sensitive microorganisms, and some species may have very specific nutritional and environmental needs. In this study, a two-phase approach was used to identify a single incubation-temperature approach that could recover most of the cleanroom microbial flora with focus on molds. Phase 1 included a growth promotion study performed in the laboratory using pharmacopeial and in-house strains, comparing different media (SDA and TSA) at single or dual incubation-temperature approaches for 5 or 6 days. Phase 2 was based on an in situ study where sampling was performed in different areas of a pharmaceutical facility, comparing the recoveries at different incubation conditions. In addition, extension studies of phase 1 and phase 2 were performed to get a better understanding of growth requirements for in-house molds. The results show that an incubation on Tryptone Soy Agar (TSA) at 25-30° for 3-4 days was able to recover most tested microorganisms in phase 1 and a large variety of microorganisms in phase 2, indicating that the single incubation-temperature is an optimal approach for the recovery of microorganisms in cleanrooms. Exceptions were noted for one strain of the species Cutibacterium acnes, a microaerophilic bacterium for which anaerobiosis and higher temperatures were needed, and two mold strains (Sistotrema brinkmannii and Stereum hirsutum), indicating that those molds required a specific media (Sabouraud Dextrose Agar, SDA) for their proliferation. The results showed that TSA incubated at single or dual incubation-temperature approach cannot compensate for the absence of SDA for some environmental molds that may be atypical in cleanrooms. Therefore, in addition to TSA, certain monitoring with SDA at e.g., cleanroom entrance points may be beneficial to recover molds with very specific nutritional requirements.

Associations of cow and farm characteristics with cow-level lameness using data from an extensive cross-sectional study across 3 structurally different dairy regions in Germany.

The aim of the present study was to evaluate the associations between milk recording data, body condition score (BCS), housing factors, management factors, and lameness in freestall-housed dairy cows in 3 structurally different regions in Germany. These regions substantially vary regarding herd size, breeds, access to pasture, farm management (family run or company owned), and percentage of organic farms. The data used was collected in a large cross-sectional study from 2016 to 2019. A total of 58,144 cows from 651 farms in 3 regions of Germany (North, East, and South) was scored for locomotion and body condition. Additionally, data on milk yield, milk composition, breed, age, as well as information on housing and management were retrieved. One mixed-logistic regression model was fitted per region to evaluate the association of the data with the target variable "lame" and to allow for a comprehensive reflection across different kinds of farming types. In all regions, undercondition (BCS lower than recommended for the lactation stage; North: odds ratio [OR] 2.15, CI 1.96-2.34; East: OR 2.66, CI 2.45-2.88; South: OR 2.45, CI 2.01-2.98) and mid-lactation stage (102-204 d in milk; North: OR 1.15, CI 1.05-1.27; East: OR 1.24, CI 1.17-1.32; South: OR 1.38, CI 1.18-1.62) were associated with higher odds for lameness, whereas overcondition (BCS higher than recommended for the lactation stage; North: OR 0.51, CI 0.44-0.60; East: OR 0.51, CI 0.48-0.54; South: OR 0.65, CI 0.54-0.77) and parity of 1 or 2 was associated with lower odds (parity 1 = North: OR 0.32, CI 0.29-0.35; East: OR 0.19, CI 0.18-0.20; South: OR 0.28, CI 0.24-0.33; parity 2 = North: OR 0.51, CI 0.47-0.46; East: OR 0.41, CI 0.39-0.44; South: OR 0.49, CI 0.42-0.57), irrespective of the regional production characteristics. Low energy-corrected milk yield was associated with higher odds for lameness in South and North (North: OR 1.16, CI 1.05-1.27; South: OR 1.43, CI 1.22-1.69). Further factors such as pasture access for cows (North: OR 0.64, CI 0.50-0.82; and South: OR 0.65, CI 0.47-0.88), milk protein content (high milk protein content = North: OR 1.34, CI 1.18-1.52; East: OR 1.17, CI 1.08-1.28; low milk protein content = North: OR 0.79, CI 0.71-0.88; East: OR 0.84, CI 0.79-0.90), and breed (lower odds for "other" [other breeds than German Simmental and German Holstein] in East [OR 0.47, CI 0.42-0.53] and lower odds both for German Holstein and "other" in South [German Holstein: OR 0.62, CI 0.43-0.90; other: OR 0.46, CI 0.34 - 0.62]) were associated with lameness in 2 regions, respectively. The risk of ketosis (higher odds in North: OR 1.11, CI 1.01-1.22) and somatic cell count (higher odds in East: increased (>39.9 cells × 1,000/mL): OR 1.10; CI 1.03-1.17; high (>198.5 cells × 1,000/mL): OR 1.08; CI 1.01-1.06) altered the odds for lameness in 1 region, respectively. Cows from organic farms had lower odds for lameness in all 3 regions (North: OR 0.18, CI 0.11-0.32; East: OR 0.39, CI 0.28-0.56; South: OR 0.45, CI 0.29-0.68). As the dairy production systems differed substantially between the different regions, the results of this study can be viewed as representative for a wide variety of loose-housed dairy systems in Europe and North America. The consistent association between low BCS and lameness in all regions aligns with the previous literature. Our study also suggests that risk factors for lameness can differ between geographically regions, potentially due to differences in which dairy production system is predominantly used and that region-specific characteristics should be taken into account in comparable future projects.

Tumour Necrosis Factor-α, Chemokines, and Leukocyte Infiltrate Are Biomarkers for Pathology in the Brains of Venezuelan Equine Encephalitis (VEEV)-Infected Mice.

Venezuelan equine encephalitis virus (VEEV) is a disease typically confined to South and Central America, whereby human disease is characterised by a transient systemic infection and occasionally severe encephalitis, which is associated with lethality. Using an established mouse model of VEEV infection, the encephalitic aspects of the disease were analysed to identify biomarkers associated with inflammation. Sequential sampling of lethally challenged mice (infected subcutaneously) confirmed a rapid onset systemic infection with subsequent spread to the brain within 24 h of the challenge. Changes in inflammatory biomarkers (TNF-α, CCL-2, and CCL-5) and CD45+ cell counts were found to correlate strongly to pathology (R>0.9) and present previously unproven biomarkers for disease severity in the model, more so than viral titre. The greatest level of pathology was observed within the olfactory bulb and midbrain/thalamus. The virus was distributed throughout the brain/encephalon, often in areas not associated with pathology. The principal component analysis identified five principal factors across two independent experiments, with the first two describing almost half of the data: (1) confirmation of a systemic Th1-biased inflammatory response to VEEV infection, and (2) a clear correlation between specific inflammation of the brain and clinical signs of disease. Targeting strongly associated biomarkers of deleterious inflammation may ameliorate or even eliminate the encephalitic syndrome of this disease.

Comparative Aspects of Ricin Toxicity by Inhalation.

The pathogenesis of ricin toxicity following inhalation has been investigated in many animal models, including the non-human primate (predominantly the rhesus macaque), pig, rabbit and rodent. The toxicity and associated pathology described in animal models are broadly similar, but variation appears to exist. This paper reviews the published literature and some of our own unpublished data and describes some of the possible reasons for this variation. Methodological variation is evident, including method of exposure, breathing parameters during exposure, aerosol characteristics, sampling protocols, ricin cultivar, purity and challenge dose and study duration. The model species and strain used represent other significant sources of variation, including differences in macro- and microscopic anatomy, cell biology and function, and immunology. Chronic pathology of ricin toxicity by inhalation, associated with sublethal challenge or lethal challenge and treatment with medical countermeasures, has received less attention in the literature. Fibrosis may follow acute lung injury in survivors. There are advantages and disadvantages to the different models of pulmonary fibrosis. To understand their potential clinical significance, these factors need to be considered when choosing a model for chronic ricin toxicity by inhalation, including species and strain susceptibility to fibrosis, time it takes for fibrosis to develop, the nature of the fibrosis (e.g., self-limiting, progressive, persistent or resolving) and ensuring that the analysis truly represents fibrosis. Understanding the variables and comparative aspects of acute and chronic ricin toxicity by inhalation is important to enable meaningful comparison of results from different studies, and for the investigation of medical countermeasures.

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).

Deep learning based on hematoxylin-eosin staining outperforms immunohistochemistry in predicting molecular subtypes of gastric adenocarcinoma.

In gastric cancer (GC), there are four molecular subclasses that indicate whether patients respond to chemotherapy or immunotherapy, according to the TCGA. In clinical practice, however, not every patient undergoes molecular testing. Many laboratories have used well-implemented in situ techniques (IHC and EBER-ISH) to determine the subclasses in their cohorts. Although multiple stains are used, we show that a staining approach is unable to correctly discriminate all subclasses. As an alternative, we trained an ensemble convolutional neuronal network using bagging that can predict the molecular subclass directly from hematoxylin-eosin histology. We also identified patients with predicted intra-tumoral heterogeneity or with features from multiple subclasses, which challenges the postulated TCGA-based decision tree for GC subtyping. In the future, deep learning may enable targeted testing for molecular subtypes and targeted therapy for a broader group of GC patients. © 2022 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Development of chlorine-induced lung injury in the anesthetized, spontaneously breathing pig.

Chlorine is a toxic industrial chemical produced in vast quantities globally, being used in a range of applications such as water purification, sanitation and industrial processes. Its use and transport cannot be restricted; exposure may occur following accidental or deliberate releases. The OPCW recently verified the use of chlorine gas against civilians in both Syria and Iraq. Chlorine inhalation produces damage to the lungs, which may result in the development of an acute lung injury, respiratory failure and death. Treatment remains an intractable problem. Our objective was to develop a clinically relevant pre-clinical model of a moderate to severe lung injury in the pig. This would enable future assessment of therapeutic drugs or interventions to be implemented in the pre-hospital phase after exposure. Due to the irritant nature of chlorine, a number of strategies for exposing terminally anesthetized pigs needed to be investigated. A number of challenges (inconsistent acute changes in respiratory parameters; early deaths), resulted in a moderate to severe lung injury not being achieved. However, most pigs developed a mild lung injury by 12 h. Further investigation is required to optimize the model and enable the assessment of therapeutic candidates. In this paper we describe the exposure strategies used and discuss the challenges encountered in establishing a model of chlorine-induced lung injury. A key aim is to assist researchers navigating the challenges of producing a clinically relevant model of higher dose chlorine exposure where animal welfare is protected by use of terminal anesthesia.

Spatial distribution and incidence of bovine neonatal pancytopenia in Bavaria, Germany.

BACKGROUND: Bovine neonatal pancytopenia (BNP) is a haemorrhagic disease of neonatal calves. BNP was first described in Germany in 2009, later on also in other European countries, and in New Zealand in 2011. The disease is characterised by spontaneous bleeding, pancytopaenia in the bone marrow, and a high case fatality ratio. The causal role of a specific bovine viral diarrhoea virus (BVDV) vaccine (PregSure®BVD, then Pfizer Animal Health, now Zoetis, Berlin, Germany) has been established over the last years, causing the production of alloantibodies in some vaccinated cattle, which in the case of pregnant cattle, are transferred to the newborn calf via the colostrum. However, striking regional differences in the incidence of the disease were observed within Germany and other countries, but as the disease was not notifiable, no representative data on the spatial distribution are available. In this study, we address the spatial distribution and incidence of BNP using the results of two representative surveys amongst cattle practitioners in Bavaria, Germany. The surveys, asking about the occurrence of BNP, were conducted in 2009 and 2010. Answers were analysed spatially by testing for clusters using space-time models. Practitioners were also asked how many cows they serve in their practice and this number was used to estimate the incidence of BNP. Furthermore, in the survey of 2010, practitioners were also asked about usage of vaccine against BVDV. RESULTS: From the results of the surveys, three clusters were identified in Bavaria. These clusters also coincided with the usage of the specific BVDV vaccine as indicated by the veterinary practices. Furthermore, the representative surveys allow the estimation of the incidence of BNP to be in the order of 4 cases per 10,000 calves at risk. CONCLUSIONS: The study is the only representative survey conducted on BNP. Despite the fact that BNP is a non-infectious disease, regional clusters were identified.

The rise of veterinary forensics.

Veterinary forensics is rapidly emerging as a distinct branch of veterinary medicine, especially because of increasing mindfulness about animal cruelty, and of the link between acts of cruelty to animals and violence toward humans. Nevertheless, the application of forensic sciences in veterinary cases lags behind its application in medical cases. Although gaps persist in veterinarians' knowledge of forensics and in how to apply this field to medicolegal cases involving animals, continued research and publication in veterinary forensics are rapidly developing the evidence base in this area. Additionally, educational opportunities in veterinary forensics are also increasing at both undergraduate and postgraduate levels. Together, these changes will continue to improve veterinarians' abilities to investigate cases involving animals. To further strengthen these investigations, veterinarians should also collaborate with the appropriate experts in different disciplines of forensic science.

Diagnosing bovine parafilariosis: utility of the cytochrome c oxidase subunit 1 gene and internal transcribed spacer region for PCR detection of Parafilaria bovicola in skin biopsies and serohemorrhagic exudates of cattle.

BACKGROUND: Parafilaria bovicola (Nematoda: Filariidae) causes cutaneous bleedings in bovine species. Flies serve as intermediate hosts. In recent years, reports on bovine parafilariosis have become more frequent, corroborating the necessity of reliable diagnostic interventions especially since no molecular or serological test has been available. We aimed to establish a polymerase chain reaction assay to detect DNA of P. bovicola in flies, skin biopsies and serohemorraghic exudates of bleeding spots. METHODS: PCRs targeting the cytochrome c oxidase subunit 1 (cox1) gene and the internal transcribed spacer region (ITS) of the ribosomal RNA gene cluster were evaluated for their diagnostic sensitivity as well as performance and specificity on biopsy and serohemorrhagic exudate samples from P. bovicola-infected cattle. RESULTS: Using serohemorrhagic exudates (n = 6), biopsies (n = 2) and flies (n = 1), the PCR targeting the cox1 gene resulted in a gel band of almost 700 bp. Cloning, sequencing, and removal of primer sequences yielded a 649-bp fragment of the P. bovicola cox1 gene. The PCR targeting the ITS region showed a band of about 1100 bp. Cloning, sequencing, and removal of primer sequences resulted in a 1083 bp stretch of the P. bovicola ITS region. Testing samples from presumably affected animals, the cox1-PCR resulted in bands with the expected size and they were all confirmed as P. bovicola by sequencing. In contrast, the ITS-PCR proved to be less sensitive and less specific and additionally amplified the ITS region of Musca domestica or buttercup DNA. When analysing for sensitivity, the cox1-PCR yielded visible bands up to 2 ng of genomic DNA, whereas the ITS-PCR produced bands up to 3 ng. In a plasmid dilution series, the minimum number of target DNA copies was 102 for the cox1-PCR and 101 in the ITS-PCR. CONCLUSIONS: The evaluated cox1-PCR enables reliable detection of P. bovicola DNA in skin biopsies and serohemorrhagic exudates. This PCR and, to a limited extent, the ITS-PCR, may help evaluate different therapeutic approaches. Furthermore, the cox1-PCR may be useful for epidemiological studies on the geographical distribution of P. bovicola. Further understanding of the epidemiology of this parasite will help develop and implement effective control strategies.

A systematic review and meta-analyses of risk factors associated with lameness in dairy cows.

BACKGROUND: Lameness in dairy cows has been an ongoing concern of great relevance to animal welfare and productivity in modern dairy production. Many studies have examined associations between various factors related to housing, management, and the individual animal and the occurrence of lameness. The objective of this systematic review was to answer the research question "what are risk factors associated with lameness in dairy cows that are housed in free stall barns or tie stall facilities". Furthermore, we performed a synthesis of current evidence on certain risk factors by means of a meta-analysis to illustrate the strength of their association with bovine lameness. RESULTS: Following pre-defined procedures and inclusion criteria in accordance with the PRISMA statement, two observers independently included 53 articles out of a pool of 1941 articles which had been retrieved by a broad literature research in a first step. 128 factors that have been associated with lameness were identified in those papers. Meta-analyses were conducted for five factors presented in six different studies: Body condition score, presence of claw overgrowth, days in milk, herd size, and parity. Results indicated that a body condition score of ≤2.5/5 is associated with increased odds of lameness. A higher risk of being lame was found for the presence of claw overgrowth, the first 120 days in milk, larger herd sizes, and increasing parity. Throughout the study, we encountered profound difficulties in retrieving data and information of sufficient quality from primary articles as well as in recovering comparable studies. CONCLUSIONS: We learned that an abundance of literature on bovine lameness exists. To adequately address a problem of this importance to both animal welfare and economic viability, solid evidence is required in the future to develop effective intervention strategies. Therefore, a consistent working definition of lameness and specific risk factors should be an option to consider.

Healed impact trauma to a Neolithic cattle frontal bone: A posthuman perspective.

Trauma associated with slaughter is identified occasionally archaeologically in the cranial remains of domesticated animals, with evidence for pole-axing occurring in Europe, especially from the Roman period onwards. The injury typically extends through the frontal bone and sinuses to penetrate the braincase, causing haemorrhage, loss of consciousness, brain damage, and death. Evidence for slaughter methods in the British Neolithic, however, is lacking. We report such evidence from a healed blunt-force impact trauma to the frontal bone of a domestic cattle skull from Beckhampton Road Neolithic long barrow, Wiltshire. The injury suggests a failed attempt at slaughter. To our knowledge, this is the first such report for domestic cattle from the British Neolithic. We contextualise this discovery, drawing on research into the role and meaning of faunal remains from Neolithic long barrows in Wiltshire. This work has been undertaken from a posthuman perspective. Thus, we demonstrate the opportunities for paleopathologists to inform and engage within posthumanist interpretative frameworks.

14-3-3σ Protein Expression in Canine Renal Cell Carcinomas.

14-3-3σ is a protein expressed in many epithelial tissues associated with essential cell functions, including cell-cycle control, apoptosis, and cytoskeletal integrity. There is a paucity of knowledge of the tumorigenesis of canine renal cell carcinomas (RCCs), and the histological origin of this tumor has not been established. This study analyzed the expression of 14-3-3σ, Ki-67, cytokeratins, and vimentin in 40 canine RCCs. Aberrant expression of 14-3-3σ was demonstrated in 15 (38%) cases and was associated with a significantly shorter survival time ( P < .002). In contrast to canine RCC, normal kidney did not express 14-3-3σ. The Ki-67 proliferation index did not show utility as a prognostic factor. The distal convoluted tubular epithelium in normal kidneys coexpressed cytokeratins and vimentin, and thus maintenance of this coexpression pattern in canine RCC suggests that most tumors arise from the distal segment of the nephron. These results suggest that 14-3-3σ is a potential negative prognostic factor and a possible therapeutic target.

Herd-level animal management factors associated with the occurrence of bovine neonatal pancytopenia in calves in a multi-country study.

Since 2007, mortality associated with a previously unreported haemorrhagic disease has been observed in young calves in several European countries. The syndrome, which has been named 'bovine neonatal pancytopenia' (BNP), is characterised by thrombocytopenia, leukocytopenia and a panmyelophthisis. A herd-level case-control study was conducted in four BNP affected countries (Belgium, France, Germany and the Netherlands) to identify herd management risk factors for BNP occurrence. Data were collected using structured face-to-face and telephone interviews of farm managers and their local veterinarians. In total, 363 case farms and 887 control farms were included in a matched multivariable conditional logistic regression analysis. Case-control status was strongly associated with the odds of herd level use of the vaccine PregSure® BVD (PregSure, Pfizer Animal Health) (matched adjusted odds ratio (OR) 107.2; 95% CI: 41.0-280.1). This was also the case for the practices of feeding calves colostrum from the calf's own dam (OR 2.0; 95% CI: 1.1-3.4) or feeding pooled colostrum (OR 4.1; 95% CI: 1.9-8.8). Given that the study had relatively high statistical power and represented a variety of cattle production and husbandry systems, it can be concluded with some confidence that no other herd level management factors are competent causes for a sufficient cause of BNP occurrence on herd level. It is suggested that genetic characteristics of the dams and BNP calves should be the focus of further investigations aimed at identifying the currently missing component causes that together with PregSure vaccination and colostrum feeding represent a sufficient cause for occurrence of BNP in calves.

OSTEOCHONDROSIS IN THE DISTAL FEMURS OF AN ADULT RETICULATED GIRAFFE (GIRAFFA CAMELOPARDALIS RETICULATA): MACROSCOPIC, RADIOLOGIC, AND HISTOLOGIC FINDINGS.

An adult male reticulated giraffe (Giraffa camelopardalis reticulata) was presented for postmortem examination. During radiologic examination of the hindlimbs, osseous cyst-like lesions were detected in both medial femoral condyles. These lesions were subsequently examined macroscopically and histologically. The gross appearance suggested a diagnosis of bilateral osteochondrosis that was confirmed with histopathologic examination. This finding has not previously been reported in giraffes. Macroscopic visualization of the major limb joints, including the femorotibial joints, is therefore encouraged in future postmortem examinations of giraffes (Giraffa camelopardalis), and further assessment of clinical significance is required.

Calf-level factors associated with bovine neonatal pancytopenia--a multi-country case-control study.

Bovine neonatal pancytopenia (BNP), a high fatality condition causing haemorrhages in calves aged less than 4 weeks, was first reported in 2007 in Germany and subsequently observed at low incidence in other European countries and New Zealand. A multi-country matched case-control study was conducted in 2011 to identify calf-level risk factors for BNP. 405 BNP cases were recruited from 330 farms in Belgium, France, Germany and the Netherlands by laboratory confirmation of farmer-reported cases. Up to four calves of similar age from the same farm were selected as controls (1154 calves). Risk factor data were collected by questionnaire. Multivariable modelling using conditional logistic regression indicated that PregSure®BVD (PregSure, Pfizer Animal Health) vaccination of the dam was strongly associated with BNP cases (adjusted matched Odds Ratio - amOR 17.8 first lactation dams; 95% confidence interval - ci 2.4, 134.4; p = 0.005), and second or more lactation PregSure-vaccinated dams were more likely to have a case than first lactation vaccinated dams (amOR 2.2 second lactation; ci 1.1, 4.3; p = 0.024; amOR 5.3 third or more lactation; ci 2.9, 9.8; p = <0.001). Feeding colostrum from other cows was strongly associated with BNP if the dam was not PregSure-vaccinated (amOR 30.5; ci 2.1, 440.5; p = 0.012), but the effect was less if the dam was PregSure-vaccinated (amOR 2.1; ci 1.1, 4.0; p = 0.024). Feeding exclusively dam's milk was a higher risk than other types of milk (amOR 3.4; ci 1.6, 7.5; p = 0.002). The population attributable fractions were 0.84 (ci 0.68, 0.92) for PregSure vaccination, 0.13 (ci 0.06, 0.19) for feeding other cows' colostrum, and 0.15 (ci 0.08, 0.22) for feeding dam's milk. No other calf-level factors were identified, suggesting that there are other important factors that are outside the scope of this study, such as genetics, which explain why BNP develops in some PregSure-colostrum-exposed calves but not in others.

Identification of essential genes for linear replication of an SCP1 composite plasmid.

The 365 023 bp linear plasmid Streptomyces coelicolor plasmid 1 (SCP1) of S. coelicolor A3(2) belongs to the model systems of actinomycetes linear plasmids. Although the plasmid was detected more than two decades ago, little functional information about replication and maintenance of the linear topology exists. Here, for the first time, is the description of a region in SCP1 that is essential for linear replication of the plasmid. The SCP1 linear replication (Slr) region contains three putative genes. Intriguingly, one of the genes, SlrC, does show homology to PPA1294 of Propionibacterium acne that possesses a conserved DNA_pol_B_2 domain, which is found in organellar and viral DNA polymerases as, for example, in the Bacillus phi29 polymerase.