Fecal microbiota composition, serum metabolomics, and markers of inflammation in dogs fed a raw meat-based diet compared to those on a kibble diet.

Introduction: Despite the potential health risks associated with feeding raw and non-traditional diets, the use of these diets in dogs is increasing, yet the health outcomes associated with these diets is not well understood. This study investigates the effect of feeding dogs a kibble or raw meat-based diets on fecal microbiota composition, serum metabolomics and inflammatory markers. Methods: Clinically healthy dogs with a history of consuming either kibble (KD, n = 27) or raw meat-based diets (RMBD, n = 28) for more than 1 year were enrolled. Dogs were fed a standardized diet of either a single brand of KD or RMBD for 28 days. Serum and fecal samples were collected for analysis of microbiota, metabolomics, and inflammatory markers. Multiple regression analysis was performed for each of the metabolites and inflammatory markers, with feed group, age and BCS included as independent variables. Results: The fecal microbiota composition differed between the KD and RMBD groups. Beta-diversity and some indices of alpha-diversity (i.e., Shannon and Simpson) were different between the two diet groups. Sixty- three serum metabolites differed between KD and RMBD-fed dogs with the majority reflecting the differences in macronutrient composition of the two diets.Fecal IAP, IgG and IgA were significantly higher in RMBD dogs compared to KD dogs, while systemic markers of inflammation, including serum c-reactive protein (CRP), galectin, secretory receptor of advanced glycation end-products (sRAGE), haptoglobin, and serum IgG were similar in dogs fed either diet. Discussion: Diet composition significantly affected fecal microbiota composition and metabolome. Although it had a potentially beneficial effect on local inflammatory markers, feeding RMBD had no impact on systemic inflammation. The influence of these changes on long term health outcomes provides an area for future study.

Heritability and variance component estimation for feed and water intake behaviors of feedlot cattle.

Feed and water intake are two important aspects of cattle production that greatly impact the profitability, efficiency, and sustainability of producers. Feed and, to a lesser degree, water intake have been studied previously; however, there is little research on their associated animal behaviors and there is a lack of standardized phenotypes for these behaviors. Feed and water intakes obtained with an Insentec system (Hokofarm Group, The Netherlands) from 830 crossbred steers were used to compute five intake behaviors for both feed and water: daily sessions (DS), intake rate (IR), session size (SS), time per session (TS), and session interval (SI). Variance components and heritabilities were estimated for each trait. Heritabilities for feed intake behaviors were 0.50 ±â€…0.12, 0.63 ±â€…0.12, 0.40 ±â€…0.13, 0.35 ±â€…0.12, and 0.60 ±â€…0.12 for DS, IR, SS, TS, and SI, respectively. Heritabilities for water intake behaviors were 0.56 ±â€…0.11, 0.88 ±â€…0.07, 0.70 ±â€…0.11, 0.54 ±â€…0.12, and 0.80 ±â€…0.10 for NS, IR, SS, TS, and SI, respectively. Daily dry matter intake (DDMI) and daily water intake (DWI) had heritabilities of 0.57 ±â€…0.11 and 0.44 ±â€…0.11. Phenotypic correlations varied between pairs of traits (-0.83 to 0.82). Genetic correlations between DDMI and feed intake behaviors were moderate to high, while genetic correlations between DWI and water intake behaviors were low to moderate. Several significant single nucleotide polymorphisms (SNP) were identified for the feed and water intake behaviors. Genes and previously reported quantitative trait loci near significant SNPs were evaluated. The results indicated that feed and water intake behaviors are influenced by genetic factors and are heritable, providing one additional route to evaluate or manipulate feed and water intake.

Feed and water intake are important aspects of cattle production to understand because they impact producer profitability and sustainability. While feed intake and, to a lesser degree, water intake have previously been studied, the associated feeding and drinking behaviors are relatively unknown and lack standardized phenotypes. Using individual animal feed and water intake records, five behaviors were evaluated for feed and water intake from crossbred feedlot steers. The behaviors evaluated were daily sessions (no./d), session size (kg), time per session (s), intake rate (g/s), and session interval (min). The impact of season (winter vs. summer) and bunk management (ad libitum vs. slick) on feeding and drinking behaviors was evaluated. Heritability and variance components were estimated for all feeding and drinking behaviors. Pairwise phenotypic correlations between behaviors were discussed. The relationship between intake and feeding or drinking behaviors was evaluated with genetic correlations. A genome-wide association study identified several significant single nucleotide polymorphisms for feeding and drinking behaviors. The results indicate that feeding and drinking behaviors are heritable and may be one additional route to evaluate feed and water intake.

Variance component estimation and genome-wide association of predicted methane production in crossbred beef steers.

Enteric methane is a potent greenhouse gas and represents an escape of energy from the ruminant digestive system. Additive genetic variation in methane production suggests that genetic selection offers an opportunity to diminish enteric methane emissions. Logistic and monetary difficulties in directly measuring methane emissions can make genetic evaluation on an indicator trait such as predicted methane production a more appealing option, and inclusion of genotyping data can result in greater genetic progress. Three predicted methane production traits were calculated for 830 crossbred steers fed in seven groups. The methane prediction equations used included mathematical models from Ellis et al. (2007), Mills et al. (2003), and IPCC (2019). Pearson correlations between the traits were all greater than 0.99, indicating that each prediction equation behaved similarly. Further, the Spearman correlations between the estimated breeding values for each trait were also 0.99, which suggests any of the predicted methane models could be used without substantially changing the ranking of the selection candidates. The heritabilities of Ellis, Mills, and IPCC predicted methane production were 0.60, 0.62, and 0.59, respectively. A genome-wide association study identified one single nucleotide polymorphism (SNP) that reached the threshold for significance for all of the traits on chromosome 7 related to oxidoreductase activity. Additionally, the SNP slightly below the significance threshold indicate genes related to collagen, intracellular microtubules, and DNA transcription may play a role in predicted methane production or its component traits.

Cattle produce methane, a greenhouse gas, as a byproduct of their digestion. It is possible to breed for animals which naturally produce less methane; however, measuring animals for methane production can be difficult or expensive and is required for effective selection. Therefore, an alternative solution is to use a mathematical model to predict methane production and select for animals with low predicted methane. The heritability of predicted methane production from each model ranged from 0.59 to 0.62. Animals were ranked nearly identical, regardless of model used. A genome-wide association study was also conducted to determine what loci may be related to predicted methane production. One significant locus was identified on chromosome 7 related to oxidoreductase activity. Other loci approaching significance showed that genes related to collagen production, intracellular microtubule binding, and DNA transcription may be related to predicted methane production. In particular, collagen turnover may have a relationship to predicted methane because it affects growth rate, which is driven by dry matter intake, which, in turn, is the primary driver of predicted methane production.

Microbial Profiling of Biltong Processing Using Culture-Dependent and Culture-Independent Microbiome Analysis.

Biltong is a South African air-dried beef product that does not have a heat lethality step, but rather relies on marinade chemistry (low pH from vinegar, ~2% salt, spices/pepper) in combination with drying at ambient temperature and low humidity to achieve microbial reduction during processing. Culture-dependent and culture-independent microbiome methodologies were used to determine the changes in the microbial community at each step during biltong processing through 8 days of drying. Culture-dependent analysis was conducted using agar-based methods to recover viable bacteria from each step in the biltong process that were identified with 16S rRNA PCR, sequencing, and BLAST searching of the NCBI nucleotide database. DNA was extracted from samples taken from the laboratory meat processing environment, biltong marinade, and beef samples at three stages of processing (post-marinade, day 4, and day 8). In all, 87 samples collected from two biltong trials with beef obtained from each of three separate meat processors (n = six trials) were amplified, sequenced with Illumina HiSeq, and evaluated with bioinformatic analysis for a culture-independent approach. Both culture-dependent and independent methodologies show a more diverse population of bacteria present on the vacuum-packaged chilled raw beef that reduces in diversity during biltong processing. The main genera present after processing were identified as Latilactobacillus sp., Lactococcus sp., and Carnobacterium sp. The high prevalence of these organisms is consistent with extended cold-storage of vacuum-packaged beef (from packers, to wholesalers, to end users), growth of psychrotrophs at refrigeration temperatures (Latilactobacillus sp., Carnobacterium sp.), and survival during biltong processing (Latilactobacillus sakei). The presence of these organisms on raw beef and their growth during conditions of beef storage appears to 'front-load' the raw beef with non-pathogenic organisms that are present at high levels leading into biltong processing. As shown in our prior study on the use of surrogate organisms, L. sakei is resistant to the biltong process (i.e., 2-log reduction), whereas Carnobacterium sp. demonstrated a 5-log reduction in the process; the recovery of either psychrotroph after biltong processing may be dependent on which was more prevalent on the raw beef. This phenomenon of psychrotrophic bloom during refrigerated storage of raw beef may result in a natural microbial suppression of mesophilic foodborne pathogens that are further reduced during biltong processing and contributes to the safety of this type of air-dried beef.

Variation in mineral types of uroliths from ferrets (Mustela putorius furo) submitted for analysis in North America, Europe, or Asia over an 8-year period.

OBJECTIVE: To compare mineral types of naturally occurring uroliths in ferrets (Mustela putorius furo) from North America, Europe, and Asia and to identify potential risk factors associated with cystine urolithiasis in ferrets. SAMPLES: 1,054 laboratory submission records of uroliths obtained from ferrets between January 1, 2010, and December 31, 2018. PROCEDURES: For this cross-sectional study, the medical records databases at 4 diagnostic laboratories were searched for records of submissions of uroliths obtained from ferrets. Data collection included submission date; ferret sex, neuter status, and age; receiving laboratory and continent; and urolith mineral type. Regression analyses were performed to identify variables associated with cystine uroliths. RESULTS: Of the 1,054 urolith submissions, 1,013 were from North America, with 92.6% (938/1,013; 95% CI, 90.8% to 94.1%) cystine uroliths, and 41 were from Europe and Asia, with only 26.8% (11/41; 95% CI, 15.7% to 41.9%) cystine uroliths. Median age was 2.0 years for ferrets with cystine urolithiasis versus 4.0 years for those with other types of uroliths. Submissions were more likely cystine uroliths for ferrets in North America versus Europe and Asia (adjusted OR [aOR], 59.5; 95% CI, 21.4 to 165.6), for ferrets that were younger (aOR, 0.67; 95% CI, 0.58 to 0.77), or for submissions in 2018 versus 2010 (aOR, 21.1; 95% CI, 5.1 to 87.9). CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that the proportion of submissions that were cystine uroliths dramatically increased in North America between 2010 and 2018. There is an urgent need to determine underlying causes and mitigate cystine urolithiasis in ferrets.

Total nitrogen is the main soil property associated with soil fungal community in karst rocky desertification regions in southwest China.

Karst rocky desertification (KRD) is a type of land deterioration, resulting in the degraded soil and a delicate ecosystem. Previous studies focused on the influence of KRD on the animals and plants, the impact of KRD on microorganisms, especially soil fungi remains to be discovered. This study reveals the change in the soil fungal community in response to KRD progression in southwest China. Illumina HiSeq was used to survey the soil fungal community. Results showed that the soil fungal community in the severe KRD (SKRD) was noticeably different from that in other KRD areas. Statistical analyses suggested that soil TN was the primary factor associated with the fungal community, followed by pH. Phylum Ascomycota was significantly abundant in non-degraded soils; whereas Basidiomycota predominated in SKRD. The ratio of Ascomycota/Basidiomycota significantly decreased along with KRD progression, which might be used as an indicator of KRD severity. Phylum Basidiomycota was sensitive to changes in all the soil properties but AP. Genus Sebacina might have the potential to promote vegetation and land restoration in KRD areas. This study fills a gap of knowledge on changes in soil fungal communities in accordance with KRD progression.

Effect of very low-protein diets supplemented with branched-chain amino acids on energy balance, plasma metabolomics and fecal microbiome of pigs.

Feeding pigs with very-low protein (VLP) diets while supplemented with limiting amino acids (AA) results in decreased growth. The objective of this study was to determine if supplementing VLP diets with branched-chain AA (BCAA) would reverse the negative effects of these diets on growth and whether this is associated with alterations in energy balance, blood metabolomics and fecal microbiota composition. Twenty-four nursery pigs were weight-matched, individually housed and allotted into following treatments (n = 8/group): control (CON), low protein (LP) and LP supplemented with BCAA (LP + BCAA) for 4 weeks. Relative to CON, pigs fed with LP had lower feed intake (FI) and body weight (BW) throughout the study, but those fed with LP + BCAA improved overall FI computed for 4 weeks, tended to increase the overall average daily gain, delayed the FI and BW depression for ~ 2 weeks and had transiently higher energy expenditure. Feeding pigs with LP + BCAA impacted the phenylalanine and protein metabolism and fatty acids synthesis pathways. Compared to CON, the LP + BCAA group had higher abundance of Paludibacteraceae and Synergistaceae and reduced populations of Streptococcaceae, Oxyphotobacteria_unclassified, Pseudomonadaceae and Shewanellaceae in their feces. Thus, supplementing VLP diets with BCAA temporarily annuls the adverse effects of these diets on growth, which is linked with alterations in energy balance and metabolic and gut microbiome profile.

Impact of Up- and Downregulation of Metabolites and Mitochondrial Content on pH and Color of the Longissimus Muscle from Normal-pH and Dark-Cutting Beef.

Limited knowledge is currently available on the biochemical basis for the development of dark-cutting beef. The objective of this research was to determine the metabolite profile and mitochondrial content differences between normal-pH and dark-cutting beef. A gas chromatography-mass spectrometer-based nontargeted metabolomic approach indicated downregulation of glycolytic metabolites, including glucose-1- and 6-phosphate and upregulation of tricarboxylic substrates such as malic and fumaric acids occurred in dark-cutting beef when compared to normal-pH beef. Neurotransmitters such as 4-aminobutyric acid and succinate semialdehyde were upregulated in dark-cutting beef than normal-pH beef. Immunohistochemistry indicated a more oxidative fiber type in dark-cutting beef than normal-pH beef. In support, the mitochondrial protein and DNA content were greater in dark-cutting beef. This increased mitochondrial content, in part, could influence oxygen consumption and myoglobin oxygenation/appearance of dark-cutting beef. The current results demonstrate that the more tricarboxylic metabolites and mitochondrial content in dark-cutting beef impact muscle pH and color.

Low Protein-High Carbohydrate Diets Alter Energy Balance, Gut Microbiota Composition and Blood Metabolomics Profile in Young Pigs.

Reducing dietary crude protein (CP) beyond a certain threshold leads to poor growth performance in pigs; however, the underlying mechanisms are not well understood. Following an adaption period, thirty-seven weaned pigs were weight matched (8.41 ± 0.14 kg), housed individually and randomly assigned into three groups with different dietary CP levels: 24% CP (CON; n = 12), 18% CP (n = 12) and 12% CP (n = 13) for 28 days. The body weight was not different between the CON and 18% CP diets, but 12% CP significantly decreased body weight after day 21. Compared to the CON, pigs fed with 12% CP decreased feed intake day 17 onwards. The 12% CP diet increased the energy expenditure during week 1 compared to the CON. The 12% CP influenced starch and sucrose, nitrogen, and branched-chain amino acids metabolism pathways. The feces of pigs fed with 12% CP were less enriched in Prevotella, but had higher relative abundance of Christensenedilaceae, Aligiphilus and Algoriphagus than CON and 18% CP. Overall, reducing dietary CP by 50%, but not by 25%, significantly influenced the physiological responses in nursery pigs. The pigs fed with low or standard protein diets had differential bacterial communities in their feces as well as serum metabolomics profile.

Characterization of water intake and water efficiency in beef cattle1,2.

In the future, water may not be as readily available due to increases in competition from a growing human population, wildlife, and other agricultural sectors, making selection for water efficiency of beef cattle increasingly important. Substantial selection emphasis has recently been placed on feed efficiency in an effort to reduce production costs, but no emphasis has been placed on making cattle more water efficient due to lack of data. Thus, the objective of this study was to calculate water efficiency metrics for cattle and evaluate their relationship to growth, feed intake (FI), and feed efficiency. Individual daily FI and water intake (WI) records were collected on 578 crossbred steers over a 70-d test period. Animals with low water intake ate less feed, had lower gains, and were more water efficient (as defined by water to gain ratio, W/G, and residual water intake, RWI). However, the amount of water consumed by animals had minimal phenotypic relationship with feed efficiency (residual feed intake [RFI], R2 = 0.1050 and feed to gain ratio (F/G) ratio R2 = 0.0726). Cattle that had low DMI consumed less water, had lower gains, had lower RFI, and had higher F/G. The level of feed consumed had minimal relationship with water efficiency. WI, W/G, RWI, and ADG had moderate heritability estimates of 0.39, 0.39, 0.37, and 0.37, respectively. High heritability estimates were observed for DMI and RFI (0.67 and 0.65, respectively). Feed to gain had a low heritability estimate of 0.16. WI had a strong positive genetic correlation with W/G (0.99) and RWI (0.88), thus selecting for decreased WI should also make cattle more water efficient. The genetic correlation between WI and ADG was 0.05; thus, selecting for low WI cattle should have little effect on growth. There is a low to moderate genetic correlation between WI and DMI (0.34). RWI has a positive genetic correlation with W/G ratio (0.89) and F/G ratio (0.42) and is negatively genetically correlated with RFI (-0.57). Water to gain and F/G had a strong positive genetic correlation (0.68). RFI has a positive genetic correlation with W/G ratio (0.37) and F/G (0.88). Minimal antagonisms seem to be present between WI and ADG, although it should be noted that standard errors were large and often not significantly different from zero due to the small sample size. However, care should be taken to ensure that unintended changes do not occur in DMI or other production traits and incorporation of WI into a selection index would likely prove to be the most effective method for selection.

Environmental effects on water intake and water intake prediction in growing beef cattle.

Water is an essential nutrient, but there are few recent studies that evaluate how much water individual beef cattle consume and how environmental factors affect an individual's water intake (WI). Most studies have focused on WI of whole pens rather than WI of individual animals. Thus, the objective of this study was to evaluate the impact of environmental parameters on individual-animal WI across different seasons and develop prediction equations to estimate WI, including within different environments and management protocols. Individual daily feed intake and WI records were collected on 579 crossbred steers for a 70-d period following a 21-d acclimation period for feed and water bunk training. Steers were fed in 5 separate groups over a 3-yr period from May 2014 to March 2017. Individual weights were collected every 14 d and weather data were retrieved from the Oklahoma Mesonet's Stillwater station. Differences in WI as a percent of body weight (WI%) were analyzed accounting for average temperature (TAVG), relative humidity (HAVG), solar radiation (SRAD), and wind speed (WSPD). Seasonal (summer vs. winter) and management differences (ad libitum vs. slick bunk) were examined. Regression analysis was utilized to generate 5 WI prediction equations (overall, summer, winter, slick, and ad libitum). There were significant (P < 0.05) differences in WI between all groups when no environmental parameters were included in the model. Although performance was more similar after accounting for all differences in weather variables, significant (P < 0.05) seasonal and feed management differences were still observed for WI%, but were less than 0.75% of steer body weight. The best linear predictors of daily WI (DWI) were dry mater intake (DMI), metabolic body weights (MWTS), TAVG, SRAD, HAVG, and WSPD. Slight differences in the coefficient of determinations for the various models were observed for the summer (0.34), winter (0.39), ad libitum (0.385), slick bunk (0.41), and overall models (0.40). Based on the moderate R2 values for the WI prediction equations, individual DWI can be predicted with reasonable accuracy based on the environmental conditions that are present, MWTS, and DMI consumed, but substantial variation exists in individual animal WI that is not accounted for by these models.

Soil pH Is the Primary Factor Correlating With Soil Microbiome in Karst Rocky Desertification Regions in the Wushan County, Chongqing, China.

Karst rocky desertification (KRD) is a process of land degradation, which causes desert-like landscapes, deconstruction of endemic biomass, and declined soil quality. The relationship of KRD progression with above-ground communities (e.g. vegetation and animal) is well-studied. Interaction of soil desertification with underground communities, such as soil microbiome, however, is vastly unknown. This study characterizes change in soil bacterial community in response to KRD progression. Soil bacterial communities were surveyed by deep sequencing of 16S amplicons. Eight soil properties, pH, soil organic matter (SOM), total and available nitrogen (TN and AN), total and available phosphorus (TP and AP), and total and available potassium (TK and AK), were measured to assess soil quality. We find that the overall soil quality decreases along with KRD progressive gradient. Soil bacterial community compositions are distinguishingly different in KRD stages. The richness and diversity in bacterial community do not significantly change with KRD progression although a slight increase in diversity was observed. A slight decrease in richness was seen in SKRD areas. Soil pH primarily correlates with bacterial community composition. We identified a core microbiome for KRD soils consisting of; Acidobacteria, Alpha-Proteobacteria, Planctomycetes, Beta-Proteobacteria, Actinobacteria, Firmicutes, Delta-Proteobacteria, Chloroflexi, Bacteroidetes, Nitrospirae, and Gemmatimonadetes in this study. Phylum Cyanobacteria is significantly abundant in non-degraded soils, suggesting that Cyanobacterial activities might be correlated to soil quality. Our results suggest that Proteobacteria are sensitive to changes in soil properties caused by the KRD progression. Alpha- and beta-Proteobacteria significantly predominated in SKRD compared to NKRD, suggesting that Proteobacteria, along with many others in the core microbiome (Acidobacteria, Actinobacteria, Firmicutes, and Nitrospirae), were active in nutrient limiting degraded soils. This study demonstrates the relationship of soil properties with bacterial community in KRD areas. Our results fill the gap of knowledge on change in soil bacterial community during KRD progression.

Test duration for water intake, ADG, and DMI in beef cattle.

Water is an essential nutrient, but the effect it has on performance generally receives little attention. There are few systems and guidelines for collection of water intake (WI) phenotypes in beef cattle, which makes large-scale research on WI a challenge. The Beef Improvement Federation has established guidelines for feed intake (FI) and ADG tests, but no guidelines exist for WI. The goal of this study was to determine the test duration necessary for collection of accurate WI phenotypes. To facilitate this goal, individual daily WI and FI records were collected on 578 crossbred steers for a total of 70 d using an Insentec system at the Oklahoma State University Willard Sparks Beef Research Unit. Steers were fed in five groups and were individually weighed every 14 d. Within each group, steers were blocked by BW (low and high) and randomly assigned to one of four pens containing approximately 30 steers per pen. Each pen provided 103.0 m2 of shade and included an Insentec system containing six feed bunks and one water bunk. Steers were fed a constant diet across groups and DMI was calculated using the average of weekly percent DM within group. Average FI and WI for each animal were computed for increasingly large test durations (7, 14, 21, 28, 35, 42, 49, 56, 63, and 70 d), and ADG was calculated using a regression formed from BW taken every 14 d (0, 14, 28, 42, 56, and 70 d). Intervals for all traits were computed starting from both the beginning (day 0) and the end of the testing period (day 70). Pearson and Spearman correlations were computed for phenotypes from each shortened test period and for the full 70-d test. Minimum test duration was determined when the Pearson correlations were greater than 0.95 for each trait. Our results indicated that minimum test duration for WI, DMI, and ADG were 35, 42, and 70 d, respectively. No comparable studies exist for WI; however, our results for FI and ADG are consistent with those in the literature. Although further testing in other populations of cattle and areas of the country should take place, our results suggest that WI phenotypes can be collected concurrently with DMI, without extending test duration, even if following procedures for decoupled intake and gain tests.

Effects of Muscle-Specific Oxidative Stress on Cytochrome c Release and Oxidation-Reduction Potential Properties.

Mitochondria play a significant role in beef color. However, the role of oxidative stress in cytochrome c release and mitochondrial degradation is not clear. The objective was to determine the effects of display time on cytochrome c content and oxidation-reduction potential (ORP) of beef longissimus lumborum (LL) and psoas major (PM) muscles. PM discolored by day 3 compared with LL. On day 0, mitochondrial content and mitochondrial oxygen consumption were greater in PM than LL. However, mitochondrial content and oxygen consumption were lower (P < 0.05) in PM than LL by day 7. Conversely, cytochrome c content in sarcoplasm was greater on days 3 and 7 for PM than LL. There were no significant differences in ORP for LL during display, but ORP increased for PM on day 3 when compared with day 0. The results suggest that muscle-specific oxidative stress can affect cytochrome c release and ORP changes.

The sheep genome illuminates biology of the rumen and lipid metabolism.

Sheep (Ovis aries) are a major source of meat, milk, and fiber in the form of wool and represent a distinct class of animals that have a specialized digestive organ, the rumen, that carries out the initial digestion of plant material. We have developed and analyzed a high-quality reference sheep genome and transcriptomes from 40 different tissues. We identified highly expressed genes encoding keratin cross-linking proteins associated with rumen evolution. We also identified genes involved in lipid metabolism that had been amplified and/or had altered tissue expression patterns. This may be in response to changes in the barrier lipids of the skin, an interaction between lipid metabolism and wool synthesis, and an increased role of volatile fatty acids in ruminants compared with nonruminant animals.

Canine intestinal histoplasmosis containing hyphal forms.

A 12-year-old intact male Miniature Schnauzer dog with chronic diarrhea that was unresponsive to empirical treatment was presented to a referring veterinarian. A laparotomy was performed, and formalin-fixed biopsies of duodenum, jejunum, and colon were sent to Oklahoma Animal Disease Diagnostic Laboratory for evaluation. Histologic examination revealed a severe, diffuse, granulomatous enteritis and colitis with intralesional yeast and hyphal forms. Grocott methenamine silver stains revealed short, aseptate hyphae co-mingled with 2-8 µm, oval to round yeast organisms consistent with Histoplasma capsulatum. The atypical presentation of both yeast and hyphal forms prompted identification of the organism. Direct sequencing of a polymerase chain reaction product from paraffin-embedded intestinal samples confirmed the presence of Ajellomyces capsulatus with a homology over 99% to several sequences in GenBank. Ajellomyces capsulatus is the holomorphic name for H. capsulatum. Therefore, the mycelial form of a dimorphic fungus such as H. capsulatum can coexist with yeast cells within lesions of histoplasmosis. Following diagnosis, the dog was treated with itraconazole for 6 months and has improved.

Transfer of Escherichia coli O157:H7 to spinach by house flies, Musca domestica (Diptera: Muscidae).

Filth flies are known mechanical vectors of pathogenic bacteria in hospital and restaurant settings, but their role as vectors for disseminating microbes to plants has not been demonstrated. Escherichia coli O157:H7 deposition by flies onto spinach was studied using molecular, microbiological, and microscopy techniques. Relative quantitative polymerase chain reaction studies showed that bacteria acquired by flies from contaminated cattle manure and deposited in regurgitation spots on leaves survived and multiplied. Scanning electron microscopy of the regurgitation spots of flies exposed to manure inoculated with E. coli suggested the multiplication of bacteria-like organisms within the spots. This finding implies that the bacteria were active and is consistent with a hypothesis that regurgitation spots serve as a nutrition source allowing E. coli O157:H7 to survive on the spinach phylloplane. E. coli O157:H7 persisted on fly body surfaces up to 13 days after exposure to acquisition sources, suggesting that fly cuticular surfaces are conducive to the growth of this pathogen. These results are consistent with the hypothesis of bioenhanced transmission of human pathogens by house flies and suggest that filth flies may affect the microbial safety of fresh produce.

Effects of aberrant estrogen on the endometrial transcriptional profile in pigs.

Estrogen prepares the uterus for implantation and is highly conserved in mammals. However, administration of estrogen to pigs earlier than normal conceptus secretion is characterized by complete embryonic mortality by days 15-18. The objective of this study was to characterize the direct effects of early estrogen administration on uterine endometrial gene expression. The study evaluated the effects of estrogen when administered early on days 9 and 10 of the estrous cycle or day 11.5 which is the normal period of conceptus estrogen secretion in pigs. Uterine endometrium was removed either Day 13 or 15. Endometrial gene expression was evaluated using the GeneChip(®) Porcine Genome Array. Fifty-five candidate genes were successfully identified as differentially expressed due to aberrant estrogen exposure, and quantitative RT-PCR mRNA expression was supportive of the array data. These data provide insight into global uterine transcriptional changes associated with estrogen disruption of the endometrium.

Dual regulation of mu opioid receptors in SK-N-SH neuroblastoma cells by morphine and interleukin-1ß: evidence for opioid-immune crosstalk.

Treatment of SK-N-SH cells with morphine and interleukin-1beta (IL-1ß) produced dual regulation of the mRNA for the human mu opioid receptor (MOR) protein. Morphine produced a decrease in the MOR mRNA while IL-1ß increased it, as assessed by real-time quantitative PCR. These data were consistent with immunocytochemical studies of treated and untreated cells. Morphine-mediated down-regulation of MOR was blocked by naltrexone and IL-1ß-induced up-regulation of MOR was blocked by interleukin-1 receptor type 1 antagonist. Immune-opioid crosstalk was examined by IL-1ß and morphine co-treatment. These data are the first to show dual regulation of MOR in neuroblastoma cells.

Endometrial caspase 1 and interleukin-18 expression during the estrous cycle and peri-implantation period of porcine pregnancy and response to early exogenous estrogen administration.

BACKGROUND: The role for endometrial secretion of cytokines during the establishment of pregnancy in a number of mammals is well established. The current study determined endometrial expression of caspase 1 (CASP1) and interleukin-18 (IL18) during the estrous cycle and early pregnancy, and following early estrogen administration, which induces conceptus loss during early development in pigs. METHODS: Gilts were hysterectomized on either D 0, 5, 10, 12, 15 and 18 of the estrous cycle, or D 10, 12, 15 or 18 of pregnancy. The abundance of endometrial CASP1 mRNA was unaffected by day of the estrous cycle, however there was a 6 and 10-fold increase in expression on D 15 and 18 of pregnancy. Endometrial expression of IL18 mRNA increased 5-fold between D 10 to 18 in cyclic and pregnant gilts. Total recoverable IL18 in uterine flushings was greater in pregnant compared to cyclic gilts on D 15 and 18.In the second experiment, mated gilts were treated with either corn oil (CO) or estrogen (E) on D 9 and 10 and hysterectomized on either D 10, 12, 13, 15 or 17 of pregnancy. The current study localizes the presence of CASP1 to the epithelial layer of the endometrium for the first time. Further, a day x treatment interaction was detected for endometrial CASP1 mRNA and protein abundance as E stimulated an earlier increase on D 13 compared to CO gilts. Although IL18 mRNA expression remained unaltered from the E treatment, protein abundance was significantly attenuated on D 15 and 18 in response to E treatment. CONCLUSIONS: Endometrial expression of CASP1 and IL18 is associated with establishment of pregnancy in pigs. Alteration of CASP1 and IL18 following premature exposure of the uterus to estrogen during early pregnancy may contribute to conceptus loss between Days 15 to 18 of pregnancy.