Small mammals as carriers of zoonotic bacteria on pig and cattle farms - Prevalence and risk of exposure in an integrative approach.

To prevent foodborne infections from pigs and cattle, the whole food chain must act to minimize the contamination of products, including biosecurity measures which prevent infections via feed and the environment in production farms. Rodents and other small mammals can be reservoirs of and key vectors for transmitting zoonotic bacteria and viruses to farm animals, through direct contact but more often through environmental contamination. In line with One Health concept, we integrated results from a sampling study of small mammals in farm environments and data from a capture-recapture experiment into a probabilistic model which quantifies the degree of environmental exposure of zoonotic bacteria by small mammals to farm premises. We investigated more than 1200 small mammals trapped in and around 38 swine and cattle farm premises in Finland in 2017/2018. Regardless of the farm type, the most common species caught were the yellow-necked mouse (Apodemus flavicollis), bank vole (Clethrionomys glareolus), and house mouse (Mus musculus). Of 554 intestine samples (each pooled from 1 to 10 individuals), 33% were positive for Campylobacter jejuni. Yersinia enterocolitica was detected in 8% of the pooled samples, on 21/38 farm premises. Findings of Salmonella and the Shiga-toxin producing Escherichia coli (STEC) were rare: the pathogens were detected in only single samples from four and six farm premises, respectively. The prevalence of Campylobacter, Salmonella, Yersinia and STEC in small mammal populations was estimated as 26%/13%, 1%/0%, 2%/3%, 1%/1%, respectively, in 2017/2018. The exposure probability within the experimental period of four weeks on farms was 17-60% for Campylobacter and 0-3% for Salmonella. The quantitative model is readily applicable to similar integrative studies. Our results indicate that small mammals increase the risk of exposure to zoonotic bacteria in animal production farms, thus increasing risks also for livestock and human health.

Evaluation of Cow-Side Meters to Determine Somatic Cell Count in Individual Cow Quarter and Bulk-Tank Milk Samples.

Intramammary infections, which cause mastitis, can increase treatment and labor costs, decrease milk production, and affect milk quality. Meters that measure quarter somatic cell count (SCC) could be used to make more informed dry cow therapy decisions. The objective of this study was to compare the RT-10 iPhone adapter (RT-10; Dairy Quality Inc., Newmarket, ON, Canada), DeLaval Cell Counter (DSCC; DeLaval, Gurnee, IL, USA), Porta Check Quick Test (PortaCheck, White City, OR, USA), California Mastitis Test (ImmuCell, Portland, ME USA), pH meter (Hanna Instruments, Smithfield, RI, USA), electrical conductivity meter (OHAUS, Parsippany, NJ, USA), and the dual laser infrared temperature thermometer (Klein Tools, Lincolnshire, IL, USA) for measuring SCC in individual Holstein mammary quarters in comparison to a reference standard, the Fourier Transform Spectrometer 600 Combi System (Combi; Bentley Instruments, Chaska, MN, USA). Meters were evaluated using 658 individual cow quarter samples and 100 bulk-tank samples to measure SCC. Individual quarter milk samples from 160 cows from four commercial dairy herds were collected just before dry off and tested within 4 h of collection. To test bulk-tank SCC, 100 bulk-tank milk samples (25 mL) were collected from UC Davis Veterinary Medicine Teaching and Research Milk Quality Lab. Meter SCC values were regressed on observed Combi SCC. Goodness of fit was then evaluated by partitioning the mean square predicted error (MSPE). For individual quarter SCC, RT-10 had the highest coefficient of determination (R2 = 0.86), lowest MSPE, and highest proportion of MSPE due to random variation (96%). Both the RT-10 and DSCC had the highest sensitivity and specificity for identifying quarter SCC above and below 200,000 cells/mL. For bulk-tank SCC, DSCC had the highest coefficient of determination (R2 = 0.45), lowest MSPE, and highest proportion of MSPE due to random variation (80%). The RT-10 and DSCC could be used to measure individual quarter SCC to determine which cows to treat at dry off potentially reducing antibiotic use.

Impact of Partially Substituting Canola Meal with Solvent-Extracted Distillers Grain with Solubles as a Protein Source on Milk Production in a Commercial Holstein Dairy Herd.

The removal of corn oil from dried distillers grains using solvent extraction allows a higher level of inclusion for solvent-extracted dried distillers grains with solubles (SDG) in rations and reduces the risk of milk fat depression in lactating Holstein dairy cows. The objective of this study was to evaluate the impact of substituting 70% of the canola meal (CM) with SDG on milk production and total mixed ration costs. A total of 1408 Holstein cows averaging 91 ± 49 days in milk were randomly enrolled to one of four dietary treatment groups: (1) primiparous control cows (PC) fed 13% CM and 0.21% urea; (2) primiparous SDG cows (PSDG) fed 4.2% CM, 8.8% SDG and 0.42% urea; (3) multiparous control cows (MC) with 13% CM and 0.21% urea; and (4) multiparous SDG cows (MSDG) with 4.1% CM, 8.6% SDG and 0.42% urea. The total mixed rations were formulated to be isonitrogenous. For the income over the feed costs from a control herd, the fed PC and MC's total mixed rations and the fed PSDG and MSDG's total mixed rations were compared. The milk yield, energy-corrected milk, milk fat yield, milk protein yield and milk protein % were lower in the PC cows compared to the PSDG cows. The MSDG cows scored lower in terms of the milk yield, milk protein yield and milk protein % and higher for the 3.5%-fat-corrected milk, milk fat yield and milk fat % compared to the MC cows. The total income, cost of dry matter and income over feed costs per cow/d were higher in the control vs. SDG simulated dairy herds. The control herd had a higher income over feed costs than the SDG herd because the average milk yield per cow/d was higher even though the SDG herd had a lower total mixed ration cost and higher milk fat production.

Genomic characterization of ESBL/AmpC-producing and high-risk clonal lineages of Escherichia coli and Klebsiella pneumoniae in imported dogs with shelter and stray background.

OBJECTIVES: Extended spectrum ß-lactamase (ESBL)- and ampicillinase C (AmpC)-carrying Enterobacteriaceae have been widely reported among companion animals. According to previous studies, dogs with a shelter or stray background might be at risk of carrying such bacteria. The aim of this study was to explore, with whole-genome sequencing (WGS), the genomic characteristics of ESBL-producing Escherichia coli and Klebsiella pneumoniae isolated from imported dogs with a stray or shelter background. METHODS: E. coli (n = 58) and K. pneumoniae (n = 2) isolates from imported dogs originating from seven countries were included. Phenotypic resistance was investigated by selective isolation and antibiotic susceptibility testing. Whole-genome sequencing was used to study the genomic characteristics and the presence of antimicrobial resistance genes (ARGs) and virulence determinants of the ESBL/AmpC-producing E. coli and K. pneumoniae isolates. RESULTS: A high diversity of different ARGs (n = 56) and sequence types (STs) (n = 32), including high-risk clonal lineages ST410 (n = 3) and ST307 (n = 1), was identified in E. coli and K. pneumoniae isolates, respectively. Genes encoding resistance to ß-lactams accounted for the majority, with the most frequent being blaCTX-M-15. Moreover, 17 (29%) E. coli isolates qualified as presumptive extraintestinal pathogenic and/or uropathogenic E. coli. CONCLUSIONS: Our results highlight the multiplicity of genetic backgrounds disseminating ESBL/AmpC-genes in the studied dogs, calling for further investigation of possible drivers responsible for the dissemination of ARGs in animal shelters and amongst stray dogs. From a public health perspective, enhanced genomic surveillance of ESBL/AmpC-producing Enterobacteriaceae in dogs is needed in Finland.

Y1 receptors modulate taste-related behavioral responsiveness in male mice to prototypical gustatory stimuli.

Mammalian taste bud cells express receptors for numerous peptides implicated elsewhere in the body in the regulation of metabolism, nutrient assimilation, and satiety. The perturbation of several peptide signaling pathways in the gustatory periphery results in changes in behavioral and/or physiological responsiveness to subsets of taste stimuli. We previously showed that Peptide YY (PYY) - which is present in both saliva and in subsets of taste cells - can affect behavioral taste responsiveness and reduce food intake and body weight. Here, we investigated the contributions of taste bud-localized receptors for PYY and the related Neuropeptide Y (NPY) on behavioral taste responsiveness. Y1R, but not Y2R, null mice show reduced responsiveness to sweet, bitter, and salty taste stimuli in brief-access taste tests; similar results were seen when wildtype mice were exposed to Y receptor antagonists in the taste stimuli. Finally, mice in which the gene encoding the NPY propeptide was deleted also showed reduced taste responsiveness to sweet and bitter taste stimuli. Collectively, these results suggest that Y1R signaling, likely through its interactions with NPY, can modulate peripheral taste responsiveness in mice.

Stable fly activity is associated with dairy management practices and seasonal weather conditions.

Stable flies (Stomoxys calcitrans) are blood-sucking insects commonly associated with cattle production systems worldwide and are known to cause severe irritation to cattle due to painful bites. Cattle react to biting stable flies with an aggregating behavior known as bunching. Bunching behavior reduces grazing or feed consumption and thus reduces cattle productivity and welfare. Cattle's fly-repelling behaviors include foot stomping, head tossing, tail switching and skin twitching. A longitudinal study was conducted in 2017 on 20 California dairies (average lactating herd size = 2,466 (SE±28.392)) during the stable fly season from April to July. The study objectives were to estimate the association between environmental factors and dairy characteristics including facility design, feed and manure management, total mixed ration (TMR) components fed to cattle, and operational pest management procedures and the outcome stable fly activity on California dairies. Stable fly activity was measured by counting stable flies on cow forelimbs (leg count) and on Alsynite traps (trap count) over the 13-week study period. Weekly leg counts were performed for cattle in lactating cow pens (31 pens from 10 study dairies) with counts made during the morning (AM) and again during the afternoon (PM). Trap counts were performed on all 20 study dairies. Data were analyzed using linear mixed models which revealed temporal variation in the average leg and trap counts with stable fly activity increasing from May to June and then decreasing to the lowest activity in July. Leg counts were higher during the afternoon compared to morning. Ambient temperatures ≤30°C and relative humidity (RH) measurements <50% were associated with higher leg and trap counts. Traps located at the periphery of study dairies had higher stable fly counts compared to traps located in the interior of the dairy. Cow pens with trees on the periphery had higher leg counts in comparison to pens away from trees. Specific TMR components were associated with both leg and trap counts. Dairies feeding by-products including almond hulls, wet distillers' grain, fruits, and vegetables had higher trap counts compared to dairies that did not feed these ingredients. At the pen level, pens with rations that contained straw had lower average leg counts compared to pens fed with rations that did not contain straw. A similar association was observed for pens with rations that contained wheat silage when ambient temperatures were ≤30°C. In contrast, pens with water added to the TMR while the RH was ≥50% had higher average leg counts compared to pens without water added to the TMR. Dairies that applied insecticides for fly control to their entire facility had lower trap counts compared to dairies that did not apply insecticides. Stable fly activity measured on California dairies using leg and trap counts varied according to the month, environmental factors, pen surroundings, trap location, TMR components, and insecticide use.

Effect of continued metabolic acidification into the first 3 days of lactation on blood calcium status in postpartum dairy cattle: A randomized controlled trial.

Although incidence of clinical hypocalcemia in postpartum dairy cows is low in US dairies, subclinical hypocalcemia after calving is common and has been associated with metabolic and infectious disease. It is widespread farm practice to feed a diet rich in anions to prepartum dairy cattle to support calcium homeostasis. However, this diet is typically discontinued at parturition, when calcium needs are still high. The objective of this trial was to determine the effects of extending metabolic acidification into the first 3 d of lactation in multiparous Holstein cows with the use of magnesium chloride (MgCl2) hexahydrate drenches on blood ionized calcium concentrations. Adult Holstein cows at a commercial dairy in their second or higher lactation, with a urine pH of 6.8 or less on the day of calving, were randomly assigned to either treatment or control groups, resulting in 13 cows in the treatment group and 14 cows in the control group. Treatment cows received 480 g of oral MgCl2 hexahydrate once daily for 3 d for continued acidification starting on the day of calving, whereas cows in the control group received no treatment. Urine pH was measured daily for 5 d, starting on the day of calving (0 DIM), to assess acidification status; blood was collected on day of calving (0 DIM), 2 DIM, and 4 DIM and analyzed for ionized calcium concentrations. Differences in blood ionized calcium and urine pH over time were compared using longitudinal data analysis. Urine pH was lower in treatment cows compared with control cows at 1, 2, and 3 DIM. Blood ionized calcium concentrations were different from baseline, taken at enrollment (0 DIM) and at 2 and 4 DIM in both treatment and control cows. However, no difference was detectable between treatment and control cows at 2 or 4 DIM with respect to blood ionized calcium concentrations. Oral supplementation with MgCl2 hexahydrate resulted in the desired acidification of urine pH in the treatment group, similar to feeding of an anionic close-up diet. Continued acidification of dairy cows until 2 DIM did not result in clinically meaningful higher blood calcium concentrations compared with controls, and further research is needed, to identify physiological reasons for this finding.

14C-Cobalamin Absorption from Endogenously Labeled Chicken Eggs Assessed in Humans Using Accelerator Mass Spectrometry.

Traditionally, the bioavailability of vitamin B-12 (B12) from in vivo labeled foods was determined by labeling the vitamin with radiocobalt (57Co, 58Co or 60Co). This required use of penetrating radioactivity and sometimes used higher doses of B12 than the physiological limit of B12 absorption. The aim of this study was to determine the bioavailability and absorbed B12 from chicken eggs endogenously labeled with 14C-B12 using accelerator mass spectrometry (AMS). 14C-B12 was injected intramuscularly into hens to produce eggs enriched in vivo with the 14C labeled vitamin. The eggs, which provided 1.4 to 2.6 µg of B12 (~1.1 kBq) per serving, were scrambled, cooked and fed to 10 human volunteers. Baseline and post-ingestion blood, urine and stool samples were collected over a one-week period and assessed for 14C-B12 content using AMS. Bioavailability ranged from 13.2 to 57.7% (mean 30.2 ± 16.4%). Difference among subjects was explained by dose of B12, with percent bioavailability from 2.6 µg only half that from 1.4 µg. The total amount of B12 absorbed was limited to 0.5-0.8 µg (mean 0.55 ± 0.19 µg B12) and was relatively unaffected by the amount consumed. The use of 14C-B12 offers the only currently available method for quantifying B12 absorption in humans, including food cobalamin absorption. An egg is confirmed as a good source of B12, supplying approximately 20% of the average adult daily requirement (RDA for adults = 2.4 µg/day).

The use of ultrasound to measure muscle depth and area in postmortem Holstein dairy calves.

Monitoring growth of neonatal dairy calves is a useful management tool to assist producers in achieving goals for reproduction and performance. The goal of this study was to examine ultrasound as an in vivo tool to quantify longissimus dorsi muscle (ribeye) linear depth and extensor carpi radialis (ECR) and semitendinosus (ST) muscle cross-sectional areas in postmortem preweaned Holstein calves. Postmortem preweaned calves (n = 137, age 13.1 d ± 15.5 SD, body weight 36.5 kg ± 7.2 SD) were obtained from two California calf ranches between April and July 2013. Two operators collected ultrasound images of the ribeye, ECR, and ST muscles using an Aloka 500V equipped with a 5-cm 7.5-MHz linear transducer. Ultrasound ribeye linear depth and ECR and ST cross-sectional areas were calculated using the Ultrasound Image Capture System. Ultrasound measurements were compared to dissected (carcass) measures. Carcass ribeye linear depth was estimated using a ruler. Dissected ECR and ST muscle cross-sectional areas were estimated by tracing muscle cross sections onto transparency paper and then photocopying, cutting out, and weighing individual paper muscle tracings. Weights of the tracings were then converted to areas using the known area of a standard 8.5 × 11 inch paper. Data were analyzed by regressing carcass estimates on observed ultrasound measurements. The coefficient of determination (R 2) indicated that ultrasound measurements were most closely associated with carcass measurements for the ST muscle (R 2 = 0.60, 0.62 for operator 1 and 2, respectively) when compared to the ribeye and ECR muscles (R 2 = 0.27, 0.41 for ribeye and 0.43, 0.32 for ECR for operator 1 and 2, respectively). The mean bias showed consistent underestimation by the ultrasound measurements when predicting carcass measurements for all three muscles and for both operators (ribeye bias = 0.15, 0.40; ECR bias = 0.95, 1.15; and ST bias = 0.73, 0.27 for operator 1 and 2, respectively). Operator contributed significantly in explaining a proportion of the variation in the regression equation for the ST muscle only, whereas calf body weight contributed significantly in explaining a proportion of the variation in the regression equation for all three muscles. The results of this study demonstrated that ultrasound measurements of the ST were the most accurate for quantifying the cross-sectional area when compared to both the ECR and ribeye in postmortem Holstein calves.

Effects of two equine digestive aid supplements on hindgut health.

Gastrointestinal disease is the number one killer of horses. Little is known about the maintenance of microbes in the equine hindgut and how to distinguish a healthy gut in a live horse. Utilization of internal and external digestibility markers and starch fermentation has been extensively studied in ruminants and is the basis for research conducted on horses. The aims of this study were to investigate the effects of two equine feed digestive aid supplements on hindgut health (HGH) as reflected in fecal pH and digestibility and to compare and validate DM digestibility measurements through the use of internal and external markers such as chromium oxide (CR), lignin (Lig), indigestible ADF (iADF), indigestible NDF (iNDF), and indigestible lignin (iLig). Nine mature Quarter horses (six geldings, three mares) were used in a crossover design, three feeding periods of 17 d (51 d total), using three treatments: control, no feed additive (CON), Smartpak (SP; Plymouth, MA), or Platinum Performance (PP; Buellton, CA). Both SP and PP contained a strain of Lactobacillus, whereas SP further supplied mannanoligosaccharides (MOS) and fructooligosaccharides (FOS) and PP supplied Saccharomyces boulardii. Within the 17-d period, horses were offered orchard grass hay and sweet cob grain and the assigned treatment daily and four CR cookies to deliver 8 g/d of CR for the last 7 d of each period. Total feces were collected from 15 to 17 d. Feed and fecal samples were dried, ground, and sent to ANALAB (Fulton, IL) for nutrient analysis. Duplicate samples of feed and feces were placed in ruminally cannulated cows for in situ determination of iADF, iNDF, and iLig to estimate digestibility. Estimated CR fecal output, CR DMI, and DM digestibilities were evaluated using the root mean square prediction error percentage of the observed mean (RMSPE), concordance correlation coefficient (CCC), and Nash-Sutcliffe efficiency methods. Marker predictive ability tests showed iADF to have the least amount of bias with the smallest RMSPE (4%), largest CCC (0.43), and the largest amount of random bias (error of dispersion = 0.45). Supplementation of PP decreased CR DM digestibility (P < 0.02). Smartpak increased fecal pH (P < 0.09), but PP had no effect on fecal pH. Therefore, SP had a beneficial effect on HGH that is believed to be due to MOS and FOS.

Mathematical absurdities in the California net energy system.

Net energy systems, such as the California Net Energy System (CNES), are useful for prediction of input:output relationships not because of fidelity to the laws of thermodynamics, but because they were designed to predict well. Unless model descriptions of input:output relationships are consistent with the laws of thermodynamics, conclusions regarding those relationships may be incorrect. Heat energy (HE) + recovered energy (RE) = ME intake (MEI) is basic to descriptions of energy utilization found in the CNES and is consistent with the laws of thermodynamics; it may be the only relationship described in the CNES consistent with the first law of thermodynamics. In the CNES, efficiencies of ME utilization for maintenance (km) and gain (kg) were estimated using ordinary least squares (OLS) equations. Efficiencies thus estimated using static linear models are often inconsistent with the biochemistry of processes underlying maintenance and gain. Reactions in support of oxidative mitochondrial metabolism are thermodynamically favorable and irreversible; these reactions yield ATP, or other high-energy phosphate bonds, used for what is generally termed maintenance. Synthesis of biomass (gain) is less thermodynamically favorable; reactions do not proceed unless coupled with hydrolysis of high-energy phosphate bonds and lie closer to equilibrium than those in support of oxidative mitochondrial metabolism. The opposite is described in the CNES (k m > k g) due to failure of partitioning of HE; insufficient HE is accounted for in maintenance. Efficiencies of ME utilization (k m and k g) as described in the CNES are variable. Further neither k m nor k g are uniformly monotonic f (ME, Mcal/kg); for ME (Mcal/kg) <0.512 or >4.26, k m are inconsistent with thermodynamically allowed values for efficiencies (>1.0); k g are a monotonically positive f (ME) concentration (Mcal/kg) for ME <3.27 Mcal/kg. For ME <1.42 Mcal/kg, k g are not in the range of thermodynamically allowed values for efficiencies (0 to 1.0). Variable efficiencies of ME utilization require that the first law may not be observed in all cases. The CNES is an excellent empirical tool for prediction of input:output relationship, but many CNES parameter estimates evaluated in this study lack consistency with biology and the laws of thermodynamics.

Measuring Liver Mitochondrial Oxygen Consumption and Proton Leak Kinetics to Estimate Mitochondrial Respiration in Holstein Dairy Cattle.

Oxygen consumption, proton motive force (PMF) and proton leak are measurements of mitochondrial respiration, or how well mitochondria are able to convert NADH and FADH into ATP. Since mitochondria are also the primary site for oxygen use and nutrient oxidation to carbon dioxide and water, how efficiently they use oxygen and produce ATP directly relates to the efficiency of nutrient metabolism, nutrient requirements of the animal, and health of the animal. The purpose of this method is to examine mitochondrial respiration, which can be used to examine the effects of different drugs, diets and environmental effects on mitochondrial metabolism. Results include oxygen consumption measured as proton dependent respiration (State 3) and proton leak dependent respiration (State 4). The ratio of State 3 / State 4 respiration is defined as respiratory control ratio (RCR) and can represent mitochondrial energetic efficiency. Mitochondrial proton leak is a process that allows dissipation of mitochondrial membrane potential (MMP) by uncoupling oxidative phosphorylation from ADP decreasing the efficiency of ATP synthesis. Oxygen and TRMP+ sensitive electrodes with mitochondrial substrates and electron transport chain inhibitors are used to measure State 3 and State 4 respiration, mitochondrial membrane PMF (or the potential to produce ATP) and proton leak. Limitations to this method are that liver tissue must be as fresh as possible and all biopsies and assays must be performed in less than 10 h. This limits the number of samples that can be collected and processed by a single person in a day to approximately 5. However, only 1 g of liver tissue is needed, so in large animals, such as dairy cattle, the amount of sample needed is small relative to liver size and there is little recovery time needed.

Genotyping and phylogenetic placement of Bacillus anthracis isolates from Finland, a country with rare anthrax cases.

BACKGROUND: Anthrax, the zoonotic disease caused by the gram-positive bacterium Bacillus anthracis, is nowadays rare in northern parts of Europe including Finland and Scandinavia. Only two minor outbreaks of anthrax in 1988 and in 2004 and one sporadic infection in 2008 have been detected in animals in Finland since the 1970's. Here, we report on two Finnish B. anthracis strains that were isolated from spleen and liver of a diseased calf related to the outbreak in 1988 (strain HKI4363/88) and from a local scrotum and testicle infection of a bull in 2008 (strain BA2968). These infections occurred in two rural Finnish regions, i.e., Ostrobothnia in western Finland and Päijänne Tavastia in southern Finland, respectively. RESULTS: The isolates were genetically characterized by PCR-based methods such as multilocus variable number of tandem repeat analysis (MLVA) and whole genome-sequence analysis (WGS). Phylogenetic comparison of the two strains HKI4363/88 and BA2968 by chromosomal single nucleotide polymorphism (SNP) analysis grouped these organisms within their relatives of the minor canonical A-branch canSNP-group A.Br.003/004 (A.Br.V770) or canonical B-branch B.Br.001/002, respectively. Strain HKI4363/88 clustered relatively closely with other members of the A.Br.003/004 lineage from Europe, South Africa, and South America. In contrast, strain BA2968 clearly constituted a new sublineage within B.Br.001/002 with its closest relative being HYO01 from South Korea. CONCLUSIONS: Our results suggest that Finland harbors both unique (autochthonous) and more widely distributed, common clades of B. anthracis. We suspect that members of the common clades such as strains HKI4363/88 have been introduced only recently by anthropogenic activities involving importation of contaminated animal products. On the other hand, autochthonous strains such as isolate BA2968 probably have an older history of their introduction into Finland as evidenced by a high number of single nucleotide variant sites in their genomes.

Energetic efficiency and the first law: the California net energy system revisited.

Models of energy utilization used in livestock production predict input:output relationships well, for all the wrong reasons. Predictive accuracy in such models is not due to fidelity to biochemistry and laws of thermodynamics, but because they were developed to predict accurately, often with little regard to biochemical consistency. Relatively static linear statistical models limit thermodynamically relevant descriptions of energy utilization, especially maintenance, in growing beef cattle and are inadequate research tools, in either ordinary least squares (OLS) or Bayesian frameworks. Metabolizable energy intake (MEI) at recovered energy (RE) = 0 (MEm) and efficiencies of ME utilization for maintenance (km) and gain (kg) were estimated for 3 independent data sets using OLS or Bayesian frameworks. Estimates of MEm differed (P < 0.05) between OLS and Bayesian estimates and were not unique, indicating model misspecification. Bayesian estimates of MEm were monotonic, positive, and nonlinear f(MEI); the range was from 6.74 to 14.8 Mcal/d. Estimates of km, the ratio of heat energy (HE) at MEI = 0 to MEm, for the 3 data sets averaged 0.590 for OLS solutions, or 0.616 for the first derivative (km, dHE/dMEI for RE = 0) of a first-order function. The first derivative (dHE/dMEI) of the OLS function was > 1.0 for MEI > 22.1 Mcal/d, counter to the laws of thermodynamics and indicated model misspecification. The Bayesian estimate of km (0.420) differed (P < 0.05) from the OLS estimate and was consistent with the efficiency of ATP synthesis. Efficiency of ME use for gain for RE > 0 (kg, OLS solutions) averaged 0.397, solutions were nonunique and single-variable OLS models were misspecified (P < 0.050) for 2 of the 3 data sets. The OLS estimate of kg differed (P < 0.05) from the estimate of kg (0.676) determined in a Bayesian framework; the latter was calculated as dRE/dMEI for RE > 0. For OLS estimates km > kg; for estimates determined in a Bayesian framework km < kg, the former is inconsistent, while the latter is consistent with the thermodynamic favorability of reactions underlying maintenance and gain. Our results show that the use of relatively fixed coefficients of maintenance in current feeding standards, mathematical descriptions of metabolic processes and concepts regarding efficiencies of energy utilization in those systems need modification to be consistent with animal biology and the laws of thermodynamics.

Phylogeography of Francisella tularensis subspecies holarctica in Finland, 1993-2011.

BACKGROUND: Finland repeatedly reports some of the highest incidences of tularaemia worldwide. To determine genetic diversity of the aetiologic agent of tularaemia, Francisella tularensis, a total of 76 samples from humans (n = 15) and animals (n = 61) were analysed. METHODS: We used CanSNPs and canINDEL hydrolysis or TaqMan MGB probes for the analyses, either directly from the clinical tissue samples (n = 21) or from bacterial isolates (n = 55). RESULTS: The genotypes of the strains were assigned to three previously described basal subspecies holarctica clades. The majority of strains (n = 67) were assigned to B.12, a clade reported to dominate in Scandinavia and Eastern Europe. A single strain was assigned to clade B.4, previously reported from North America, Europe and China. The remaining strains (n = 8) were members of clade B.6. Importantly, new diversity was discovered in clade B.6. We describe two newly designed TaqMan MGB probe assays for this new B.6 subclade B.70, and its previously identified sister clade B.11, a clade dominantly found in Western Europe. CONCLUSIONS: The high genetic diversity of F. tularensis subspecies holarctica present in Finland is consistent with previous findings in Sweden. The results suggest a northern and southern division of the B.6 subclade B.10, where B.11 predominates in Western and Central Europe and B.70 is found in Fennoscandia. Further research is required to define whether the vast diversity of genotypes found is related to different habitats or reservoir species, their different postglacial immigration routes to Fennoscandia, or dynamics of the reservoir species.

Experimental Infection of voles with Francisella tularensis indicates their amplification role in tularemia outbreaks.

Tularemia outbreaks in humans have been linked to fluctuations in rodent population density, but the mode of bacterial maintenance in nature is unclear. Here we report on an experiment to investigate the pathogenesis of Francisella tularensis infection in wild rodents, and thereby assess their potential to spread the bacterium. We infected 20 field voles (Microtus agrestis) and 12 bank voles (Myodes glareolus) with a strain of F. tularensis ssp. holarctica isolated from a human patient. Upon euthanasia or death, voles were necropsied and specimens collected for histological assessment and identification of bacteria by immunohistology and PCR. Bacterial excretion and a rapid lethal clinical course with pathological changes consistent with bacteremia and tissue necrosis were observed in infected animals. The results support a role for voles as an amplification host of F. tularensis, as excreta and, in particular, carcasses with high bacterial burden could serve as a source for environmental contamination.

Computer modeling of obesity links theoretical energetic measures with experimental measures of fuel use for lean and obese men.

The goal of this research was to use a computational model of human metabolism to predict energy metabolism for lean and obese men. The model is composed of 6 state variables representing amino acids, muscle protein, visceral protein, glucose, triglycerides, and fatty acids (FAs). Differential equations represent carbohydrate, amino acid, and FA uptake and output by tissues based on ATP creation and use for both lean and obese men. Model parameterization is based on data from previous studies. Results from sensitivity analyses indicate that model predictions of resting energy expenditure (REE) and respiratory quotient (RQ) are dependent on FA and glucose oxidation rates with the highest sensitivity coefficients (0.6, 0.8 and 0.43, 0.15, respectively, for lean and obese models). Metabolizable energy (ME) is influenced by ingested energy intake with a sensitivity coefficient of 0.98, and a phosphate-to-oxygen ratio by FA oxidation rate and amino acid oxidation rate (0.32, 0.24 and 0.55, 0.65 for lean and obese models, respectively). Simulations of previously published studies showed that the model is able to predict ME ranging from 6.6 to 9.3 with 0% differences between published and model values, and RQ ranging from 0.79 to 0.86 with 1% differences between published and model values. REEs >7 MJ/d are predicted with 6% differences between published and model values. Glucose oxidation increases by ∼0.59 mol/d, RQ increases by 0.03, REE increases by 2 MJ/d, and heat production increases by 1.8 MJ/d in the obese model compared with lean model simulations. Increased FA oxidation results in higher changes in RQ and lower relative changes in REE. These results suggest that because fat mass is directly related to REE and rate of FA oxidation, body fat content could be used as a predictor of RQ.

Detection of Francisella tularensis in voles in Finland.

Francisella tularensis is a highly virulent intracellular bacterium causing the zoonotic disease tularemia. It recurrently causes human and animal outbreaks in northern Europe, including Finland. Although F. tularensis infects several mammal species, only rodents and lagomorphs seem to have importance in its ecology. Peak densities of rodent populations may trigger tularemia outbreaks in humans; however, it is still unclear to which extent rodents or other small mammals maintain F. tularensis in nature. The main objective of this study was to obtain information about the occurrence of F. tularensis in small mammals in Finland. We snap-trapped 547 wild small mammals representing 11 species at 14 locations around Finland during 6 years and screened them for the presence of F. tularensis DNA using PCR analysis. High copy number of F. tularensis-specific DNA was detected in tissue samples of five field voles (Microtus agrestis) originating from one location and 2 years. According to DNA sequences of the bacterial 23S ribosomal RNA gene amplified from F. tularensis-infected voles, the infecting agent belongs to the subspecies holarctica. To find out the optimal tissue for tularemia screening in voles, we compared the amounts of F. tularensis DNA in lungs, liver, spleen, and kidney of the infected animals. F. tularensis DNA was detectable in high levels in all four organs except for one animal, whose kidney was F. tularensis DNA-negative. Thus, at least liver, lung, and spleen seem suitable for F. tularensis screening in voles. Thus, liver, lung, and spleen all seem suitable for F. tularensis screening in voles. In conclusion, field voles can be heavily infected with F. tularensis subsp. holarctica and thus potentially serve as the source of infection in humans and other mammals.

[Botulinum toxin as a biological weapon]. / Botulinumtoksiini biouhka-agenssina.

Botulism is caused by botulinum neurotoxin produced by the bacterium Clostridium botulinum. It is a flaccid paralysis in which consciousness and nociception are preserved. Natural botulism typically results from ingestion of inadequately heated or unheated vacuum-packed foods. In addition, botulinum toxin is one of the most feared biological weapons. In the diagnosis and treatment of botulism early suspicion is essential. Several coinciding or local clusters without a typical connecting source, or an uncommon type of toxin may indicate an intentionally caused epidemic.