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Background Payment programs based on milk quality (PPBMQ) are used in several countries around the world as an incentive to improve milk quality. One of the principal milk parameters used in such programs is the bulk tank somatic cell count (BTSCC). In this study, using data from an average of 37,000 farms per month in Brazil where milk was analyzed, BTSCC data were divided into different payment classes based on milk quality. Then, descriptive and graphical analyses were performed. The probability of a change to a worse payment class was calculated, future BTSCC values were predicted using time series models, and financial losses due to the failure to reach the maximum bonus for the payment based on milk quality were simulated. Results In Brazil, the mean BTSCC has remained high in recent years, without a tendency to improve. The probability of changing to a worse payment class was strongly affected by both the BTSCC average and BTSCC standard deviation for classes 1 and 2 (1000–200,000 and 201,000–400,000 cells/mL, respectively) and only by the BTSCC average for classes 3 and 4 (401,000–500,000 and 501,000–800,000 cells/mL, respectively). The time series models indicated that at some point in the year, farms would not remain in their current class and would accrue financial losses due to payments based on milk quality. Conclusion The BTSCC for Brazilian dairy farms has not recently improved. The probability of a class change to a worse class is a metric that can aid in decision-making and stimulate farmers to improve milk quality. A time series model can be used to predict the future value of the BTSCC, making it possible to estimate financial losses and to show, moreover, that financial losses occur in all classes of the PPBMQ because the farmers do not remain in the best payment class in all months.

Data in this article depict patterns of methylation in lung tissues obtained from the offspring of B6129SF1/J dams and 129S1/SvImJ sires exposed in utero to benzo[ a ]pyrene (BaP) or dibenzo[ def,p ]chrysene (DBC) as compared to non-exposed offspring. Genome-wide methylation of lung tumors in adult offspring was determined using methylated DNA immunoprecipitation (MeDIP) with the NimbleGen mouse DNA methylation CpG island array. This data article refers to the research article “DNA methylation in lung tissues of mouse offspring exposed in utero to polycyclic aromatic hydrocarbons,” [1] in which comprehensive data interpretation and analysis are provided.

Cheddar cheese was made using control culture (Lactococcus lactis subsp. lactis), or with control culture plus a galactose-metabolising (Gal+) or galactose-non-metabolising (Gal−) Streptococcus thermophilus adjunct; for each culture type, the pH at whey drainage was either low (pH 6.15) or high (pH 6.45). S. thermophilus affected the levels of residual lactose and galactose, and the volatile compound profile and sensory properties of the mature cheese (270 d) to an extent dependent on the drain pH and phenotype (Gal+ or Gal−). For all culture systems, reducing drain pH resulted in lower levels of moisture and lactic acid, a higher concentration of free amino acids, and higher firmness. The results indicate that S. thermophilus may be used to diversify the sensory properties of Cheddar cheese, for example from a fruity buttery odour and creamy flavour to a more acid taste, rancid odour, and a sweaty cheese flavour at high drain pH.

BackgroundOver the last 10 years, the discovery of individual, or candidate, genes that influence economically important traits has revolutionised how the dairy industry selects cows for breeding programmes. One such gene, the diacylglycerol O-acyltransferase 1 (DGAT1) gene is significant in dairy production as it is associated with the synthesis of fat in milk, and polymorphisms within the gene can be used for selection purposes. In this study, the variants (genotypes) of the K232A polymorphism of the DGAT1 gene, within a dual-purpose dairy herd, bred for both dairy and beef production, were determined, and compared to the Economic Breeding Index (EBI) and EBI sub-indexes to ascertain if any statistically significant relationships existed. MethodsDNA was extracted from hair samples, and the genotypes of the DGAT1 gene were determined using a combination of conventional PCR, restriction fragment length polymorphism (RFLP), gel electrophoresis, DNA sequencing and novel real-time PCR assays. ResultsThe three genotypes within the herd were: KK (n = 5), AA (n = 10), KA (n = 8). KK denoted a homologous Lysine allele, AA denoted a homologous Alanine allele, and KA denoted a heterologous Lysine allele. There was no association found between the milk production traits and the DGAT1 K232A genotype (P = 0.178). However, there was a significant association found between the K232A polymorphism and other EBI traits including ‘Beef’ and ‘Management’ which had P values of 0.035 and 0.023 respectively. The AA genotype was associated with higher beef traits (higher carcass values) and lower management traits (reduced milking time and good temperament). The KK genotype was associated with higher management traits and lower beef traits. These results were not always consistent with those in the literature. ConclusionsThis study showed that animals with the A allele of the DGAT1 gene polymorphism may be better suited for dual-purpose dairy herds. It is also possible that different breeding goals and farm environments may influence the results suggesting that these types of studies should be conducted on a variety of herds and include larger samples sizes.

The TORC2 gene is a member of the transducer of the regulated cyclic adenosine monophosphate (cAMP) response element binding protein gene family, which plays a key role in metabolism and adipogenesis. In the present study, we confirmed the role of TORC2 in bovine preadipocyte proliferation through cell cycle staining flow cytometry, cell counting assay, 5-ethynyl-2&prime, deoxyuridine staining (EdU), and mRNA and protein expression analysis of proliferation-related marker genes. In addition, Oil red O staining analysis, immunofluorescence of adiponectin, mRNA and protein level expression of lipid related marker genes confirmed the role of TORC2 in the regulation of bovine adipocyte differentiation. Furthermore, the transcription start site and sub-cellular localization of the TORC2 gene was identified in bovine adipocytes. To investigate the underlying regulatory mechanism of the bovine TORC2, we cloned a 1990 bp of the 5' untranslated region (5&prime, UTR) promoter region into a luciferase reporter vector and seven vector fragments were constructed through serial deletion of the 5&prime, UTR flanking region. The core promoter region of the TORC2 gene was identified at location &minus, 314 to &minus, 69 bp upstream of the transcription start site. Based on the results of the transcriptional activities of the promoter vector fragments, luciferase activities of mutated fragments and siRNAs interference, four transcription factors (CCAAT/enhancer-binding protein C/BEP, X-box binding protein 1 XBP1, Insulinoma-associated 1 INSM1, and Zinc finger protein 263 ZNF263) were identified as the transcriptional regulators of TORC2 gene. These findings were further confirmed through Electrophoretic Mobility Shift Assay (EMSA) within nuclear extracts of bovine adipocytes. Furthermore, we also identified that C/EBP, XBP1, INSM1 and ZNF263 regulate TORC2 gene as activators in the promoter region. We can conclude that TORC2 gene is potentially a positive regulator of adipogenesis. These findings will not only provide an insight for the improvement of intramuscular fat in cattle, but will enhance our understanding regarding therapeutic intervention of metabolic syndrome and obesity in public health as well.

Zika virus (ZIKV) is an arboviral infection that has been shown to be sexually transmitted. The study outlined herein aims to determine if accessory sex glands and epididymal epithelial cells are sources of viral persistence in subacute and chronic ZIKV infection, and if infection of these organs is important in sexual transmission during long-term (chronic) infection. Male interferon type I receptor knockout (Ifnar−/−) mice were challenged with ZIKV and reproductive tissues were harvested 14 and 35 days post infection (DPI) for inoculation studies and 14, 35 and 70 DPI for histopathology. Artificial insemination fluid derived from epididymal flush and seminal plasma from the prostate and seminal vesicle was obtained from ZIKV inoculated and sham-infected males. Naïve interferon type I and II receptor knockout (AG129) female mice were pre-treated with progesterone and inoculated intravaginally with artificial insemination fluid from ZIKV-infected males. ZIKV RNA was detected in the artificial insemination fluid generated from epididymal flush or seminal plasma from ZIKV infected males at 14 and 35 DPI. ZIKV antigens were only detected in seminiferous tubules at 14 DPI. Epididymal epithelial cells did not show ZIKV antigen immunoreactivity at 14, 35 or 70 DPI. Severe fibrosing orchitis (end stage orchitis) was observed at 35 and 70 DPI. Mild inflammation and peri-tubular fibrosis were observed in the epididymis following clearance of virus. Viral RNA was not detected by PCR in whole blood samples of females from any intravaginal experimental group and only detected in 20% of subcutaneously inoculated animals (derived from 1 experimentally infected male) at 35 DPI. While ZIKV RNA and antigens can be detected in the male reproductive tract at 14 DPI and RNA can also be detected at 35 DPI, intravaginal inoculation of artificial insemination fluid from these time-points failed to result in viremia in naïve females inoculated intravaginally. These studies support the hypothesis that epididymal epithelial cells are critical to sexual transmission in immunocompromised mice. Additionally, acute but not chronic male reproductive tract infection with ZIKV results in infectious virus capable of being sexually transmitted in mice.

A seven-year-old spayed female Labrador retriever presented for necropsy following an acute history of thrombocytopenia, anemia, leukocytosis and abdominal effusion. A 2 × 3 × 10 cm, cylindrical to tubular, mottled red-to-tan mass extended from the caudal pelvic cavity caudally and ventrally under the dermis along the caudal aspect of the left pelvic limb adjacent to the semimembranosus and semitendinosus musculature. Histologic examination of the mass revealed a singular central lumen lined by urothelium that multifocally transitioned into non-keratinizing, stratified squamous epithelium associated with few hair follicles and sweat glands. The lumen was surrounded by a dense collagenous stroma containing numerous, variably sized blood vessels. The combination of lesions was consistent with a diagnosis of incomplete urethral duplication associated with a dermoid cyst and vascular hamartoma. To the authors’ knowledge, this is the first report of an incomplete urethral duplication associated with a dermoid cyst within a vascular hamartoma.

In 2008, a consortium led by the Agricultural Research Service (ARS) and the National Institute for Food and Agriculture (NIFA) published the “Blueprint for USDA Efforts in Agricultural Animal Genomics 2008–2017,” which served as a guiding document for research and funding in animal genomics. In the decade that followed, many of the goals set forth in the blueprint were accomplished. However, several other goals require further research. In addition, new topics not covered in the original blueprint, which are the result of emerging technologies, require exploration. To develop a new, updated blueprint, ARS and NIFA, along with scientists in the animal genomics field, convened a workshop titled “Genome to Phenome: A USDA Blueprint for Improving Animal Production” in November 2017, and these discussions were used to develop new goals for the next decade. Like the previous blueprint, these goals are grouped into the broad categories “Science to Practice,” “Discovery Science,” and “Infrastructure.” New goals for characterizing the microbiome, enhancing the use of gene editing and other biotechnologies, and preserving genetic diversity are included in the new blueprint, along with updated goals within many genome research topics described in the previous blueprint. The updated blueprint that follows describes the vision, current state of the art, the research needed to advance the field, expected deliverables, and partnerships needed for each animal genomics research topic. Accomplishment of the goals described in the blueprint will significantly increase the ability to meet the demands for animal products by an increasing world population within the next decade.

Development of antiviral drug resistance is a continuous concern for viruses with high mutation rates such as influenza. The use of antiviral drugs targeting host proteins required for viral replication is less likely to result in the selection of resistant viruses than treating with direct-acting antivirals. The iminosugar UV-4B is a host-targeted glucomimetic that inhibits endoplasmic reticulum α-glucosidase I and II enzymes resulting in improper glycosylation and misfolding of viral glycoproteins. UV-4B has broad-spectrum antiviral activity against diverse viruses including dengue and influenza. To examine the ability of influenza virus to develop resistance against UV-4B, mouse-adapted influenza virus was passaged in mice in the presence or absence of UV-4B and virus isolated from lungs was used to infect the next cohort of mice, for five successive passages. Deep sequencing was performed to identify changes in the viral genome during passaging in the presence or absence of UV-4B. Relatively few minor variants were identified within each virus and the ratio of nonsynonymous to synonymous (dN/dS) substitutions of minor variants confirmed no apparent positive selection following sustained exposure to UV-4B. Three substitutions (one synonymous in PB2, one nonsynonymous in M and PA each) were specifically enriched (>3%) in UV-4B-treated groups at passage five. Recombinant viruses containing each individual or combinations of these nonsynonymous mutations remained sensitive to UV-4B treatment in mice. Overall, these data provide evidence that there is a high genetic barrier to the generation and selection of escape mutants following exposure to host-targeted iminosugar antivirals.

It has previously been shown that young, cycling ovarian transplantation in aged female mice increased the general health and life span in regard to their post-reproductive health. It has further been hypothesized that this enhancement of health is directly influenced by the ovarian somatic cells. To address this hypothesis, transplants of young germ cell depleted and germ cell containing ovaries were performed on female mice. The purpose of this study is to continue to discern the reproductive influence on aging health, specifically in the area of immunological well-being. Control group mice were separated by age and treatment mice were subsequently age matched to receive either germ cell depleted or germ cell containing ovary transplantations. All groups underwent various health span assays including immunoassays using flow cytometric analysis to determine T-cell subset alterations. Data collected from the immunoassays were analyzed with two-factor ANOVA and a Tukey-Kramer post-hoc test to determine any difference between groups. Ratio differences and trends were analyzed between the groups and between central naïve and central memory T-cells. Results showed that the group having received germ cell depleted ovaries displayed a shift in the central naïve to central memory ratio to an increased central naïve population, thus indicating an improvement of immunological function as compared to the other test groups. This may indicate that the ovarian somatic cell participates heavily in the regulation of age-associated health.

Dietary interventions with probiotic lactic acid bacteria (LAB) and prebiotics, complex dietary fibers that promote LAB growth, may favorably shift the gut microbiome to reduce colorectal cancer risk. Our primary hypothesis was that the LAB strains NCFM Lactobacillus acidophilus and HNO 19 Bifidobacterium lactis would thrive in the presence of agave, green banana, black raspberry, baobab fruit, or pomegranate peel whole food powders by fermenting their oligosaccharide (OS) components into lactic acid end products. LAB strains were cultured in media with no carbohydrate, purified OS, or one of the whole food powders. LAB strains cultured with agave appeared to grow better compared to the purified OS treatment as determined by the optical density measurement (p < 0.0001). Also, the medium pH for L. acidophilus cultured with agave or black raspberry decreased significantly more than with purified OS (p < 0.0001 ), indicating a greater rate of fermentation. For B. lactis cultured with agave, black raspberry, or baobab fruit the medium pH was not significantly different from the purified OS, suggesting that these foods are fermented at least as well as the purified OS. However, the effect of green banana, black raspberry, or pomegranate peel on bacteria growth could not be determined because the culture medium was too opaque for measurements. The medium pH of L. acidophilus cultured with baobab fruit was not significantly different than the no carbohydrate control. Optical density measurements suggested that baobab fruit did not support growth of L. acidophilus, though B. lactis grew in the presence of baobab as well as the purified OS. Lastly, neither green banana nor pomegranate peel appeared to be utilized by LAB strains as culture media pH was not changed. In conclusion, results of this study suggest that agave and black raspberry powders support lactic acid bacteria fermentation, while green banana and pomegranate peel did not under the culture conditions employed in this study. Additionally, the fermentation of baobab powder appeared to be dependent on the probiotic strain. Of the bioactive food powders tested, black raspberry appears to be the most promising candidate for future synbiotic colorectal cancer dietary intervention studies.

Mastitis, an infection of the mammary gland, is the most common and expensive animal health problem for the dairy industry and affects every dairy farm to some degree. This disease complex is painful for dairy cows, increases the on-farm use of antibiotics, presents a threat to milk quality and is a waste of time, money and milk production. Each year, the dairy industry loses as much as a billion dollars to mastitis. Many cows will experience mastitis at least once during a lactation cycle and some animals will develop recurring mastitis episodes in a single mammary quarter. These mastitic quarters can be difficult to manage during the lactation cycle. Cessation of production in the quarter while continuing to milk the other three can be a beneficial management decision in this scenario. However, the current methods available for cessation of lactation in a single quarter are limited. This study investigated the use of casein hydrolysate as a non-antibiotic option for causing cessation of lactation in a quarter. From this preliminary study we were able to apply our results to another aspect of mastitis prevention, which is the routine use of intramammary antibiotics at the end of the lactation cycle. This management practice is known as dry treatment and is a standard practice in the dairy industry with many years of proven efficacy against clearing infections present at the end of the lactation cycle. Increasing pressure from consumers to decrease antibiotic use in food production animals has caused this practice to come under scrutiny. This secondary study investigated the use of casein hydrolysates as a non-antibiotic alternative to standard antibiotic dry cow treatment. Overall, these studies demonstrated that casein hydrolysate has some efficacy in inducing mammary involution of a single quarter mid-lactation and also potentially as an alternative dry cow treatment. None of the animals treated in these studies displayed any symptoms of pain or discomfort, and all treated quarters resumed milk production after the next calving. Additionally, all antimicrobial milk tests on treated animals were negative. Casein hydrolysates may be a useful management tool for milk quality and animal health within the dairy industry.

Susceptibility and/or resistance to aflatoxin B1 (AFB1) is a threshold trait governed principally by glutathione S transferase (GST)-mediated detoxification. In poultry, domesticated turkeys are highly sensitive to AFB1, most likely due to dysfunction in hepatic GSTs. In contrast, wild turkeys are comparatively resistant to aflatoxicosis due to the presence of functional hepatic GSTAs and other possible physiological and immunological interactions. The underlying genetic basis for the disparate GST function in turkeys is unknown as are the broader molecular interactions that control the systemic response. This study quantifies the effects of dietary AFB1 on gene expression in the turkey spleen, specifically contrasting genetically distinct domesticated (DT, susceptible) and Eastern wild (EW, resistant) birds. Male turkey poults were subjected to a short-term AFB1 treatment protocol with feed supplemented with 320 ppb AFB1 beginning on day 15 of age and continuing for 14 days. Spleen tissues were harvested and subjected to deep RNA sequencing and transcriptome analysis. Analysis of differential gene expression found the effects of AFB1 treatment on the spleen transcriptomes considerably more prominent in the DT birds compared to EW. However, expression of the differentially expressed genes (DEGs) was directionally biased, with the majority showing higher expression in EW (i.e., down-regulation in DT). Significantly altered pathways included FXR/RXR and LXR/RXR activation, coagulation system, prothrombin activation, acute phase response, and atherosclerosis signaling. Differential extra-hepatic expression of acute phase protein genes was confirmed by quantitative real time PCR (qRT-PCR) in the original experiment and additional turkey lines. Results demonstrate that wild turkeys possess a capacity to more effectively respond to AFB1 exposure.

Crystalline silica (cSiO2) is a widely recognized environmental trigger of autoimmune disease. In the lupus-prone female NZBWF1 mouse, airway exposure to cSiO2 triggers pulmonary ectopic lymphoid neogenesis, systemic autoantibody elevation, and glomerulonephritis. Here we tested the hypothesis that upregulation of adaptive immune function genes in the lung precedes cSiO2-triggering of autoimmune disease in this model. The study include three groups of mice, as follows: (1) necropsied 1 d after a single intranasal instillation of 1 mg cSiO2 or vehicle, (2) necropsied 1 d after four weekly single instillations of 1 mg cSiO2 or vehicle, or (3) necropsied 1, 5, 9, or 13 weeks after four weekly single instillations of 1 mg cSiO2 or vehicle. NanoString nCounter analysis revealed modest transcriptional changes associated with innate and adaptive immune response as early as 1 d after a single cSiO2 instillation. These responses were greatly expanded after four weekly cSiO2 instillations. Concurrent with ectopic lymphoid neogenesis, dramatic increases in mRNAs associated with chemokine release, cytokine production, sustained interferon activity, complement activation, and adhesion molecules were observed. As disease progressed, expression of these genes persisted and was further amplified. Consistent with autoimmune pathogenesis, the time between 5 and 9 weeks post-instillation reflected an important transition period where considerable immune gene upregulation in the lung was observed. Upon termination of the chronic study (13 weeks), cSiO2-induced changes in transcriptome signatures were similarly robust in kidney as compared to the lung, but more modest in spleen. Transcriptomic signatures in lung and kidney were indicative of infiltration and/or expansion of neutrophils, macrophages, dendritic cells, B cells, and T cells that corresponded with accelerated autoimmune pathogenesis. Taken together, airway exposure to cSiO2 elicited aberrant mRNA signatures for both innate and adaptive immunity that were consistent with establishment of the lung as the central autoimmune nexus for launching systemic autoimmunity and ultimately, kidney injury.

This issue has two articles. The first focuses on taxes, and financial planning. The second article discusses the Collegiate Dairy Challenge.

Colorectal cancer (CRC) is the second leading cause of cancer death in the United States. Modification of the gut microbiome offers potential for CRC prevention.

To address the unmet needs for human polyclonal antibodies both as therapeutics and diagnostic reagents, building upon our previously established transchromosomic (Tc) cattle platform, we report herein the development of a Tc goat system expressing human polyclonal antibodies in their sera. In the Tc goat system, a human artificial chromosome (HAC) comprising the entire human immunoglobulin (Ig) gene repertoire in the germline configuration was introduced into the genetic makeup of the domestic goat. We achieved this by transferring the HAC into goat fetal fibroblast cells followed by somatic cell nuclear transfer for Tc goat production. Gene and protein expression analyses in the peripheral blood mononuclear cells (PBMC) and the sera, respectively, of Tc caprine demonstrated the successful expression of human Ig genes and antibodies. Furthermore, immunization of Tc caprine with inactivated influenza A (H7N9) viruses followed by H7N9 Hemagglutinin 1 (HA1) boosting elicited human antibodies with high neutralizing activities against H7N9 viruses in vitro. As a small ungulate, Tc caprine offers the advantages of low cost and quick establishment of herds, therefore complementing the Tc cattle platform in responses to a range of medical needs and diagnostic applications where small volumes of human antibody products are needed.

Zika virus (ZIKV) infection can cause severe congenital diseases, such as microcephaly, ocular defects and arthrogryposis in fetuses, and Guillain&ndash, Barr&eacute, syndrome in adults. Efficacious therapeutic treatments for infected patients, as well as prophylactic treatments to prevent new infections are needed for combating ZIKV infection. Here, we report that ZIKV-specific human polyclonal antibodies (SAB-155), elicited in transchromosomal bovine (TcB), provide significant protection from infection by ZIKV in STAT2 knockout (KO) golden Syrian hamsters both prophylactically and therapeutically. These antibodies also prevent testicular lesions in this hamster model. Our data indicate that antibody-mediated immunotherapy is effective in treating ZIKV infection. Because suitable quantities of highly potent human polyclonal antibodies can be quickly produced from the TcB system against ZIKV and have demonstrated therapeutic efficacy in a small animal model, they have the potential as an effective countermeasure against ZIKV infection.

Currently, there are no licensed human vaccines or antivirals for treatment of or prevention from infection with encephalitic alphaviruses. Because epidemics are sporadic and unpredictable, and endemic disease is common but rarely diagnosed, it is difficult to identify all populations requiring vaccination; thus, an effective post-exposure treatment method is needed to interrupt ongoing outbreaks. To address this public health need, we have continued development of ML336 to deliver a molecule with prophylactic and therapeutic potential that could be relevant for use in natural epidemics or deliberate release scenario for Venezuelan equine encephalitis virus (VEEV). We report findings from in vitro assessments of four analogs of ML336, and in vivo screening of three of these new derivatives, BDGR-4, BDGR-69 and BDGR-70. The optimal dosing for maximal protection was observed at 12.5 mg/kg/day, twice daily for 8 days. BDGR-4 was tested further for prophylactic and therapeutic efficacy in mice challenged with VEEV Trinidad Donkey (TrD). Mice challenged with VEEV TrD showed 100% and 90% protection from lethal disease when treated at 24 and 48 h post-infection, respectively. We also measured 90% protection for BDGR-4 in mice challenged with Eastern equine encephalitis virus. In additional assessments of BDGR-4 in mice alone, we observed no appreciable toxicity as evaluated by clinical chemistry indicators up to a dose of 25 mg/kg/day over 4 days. In these same mice, we observed no induction of interferon. Lastly, the resistance of VEEV to BDGR-4 was evaluated by next-generation sequencing which revealed specific mutations in nsP4, the viral polymerase.

The nearly-ubiquitous food and feed-borne mycotoxin aflatoxin B1 (AFB1) is carcinogenic and mutagenic, posing a food safety threat to humans and animals. One of the most susceptible animal species known and thus a good model for characterizing toxicological pathways, is the domesticated turkey (DT), a condition likely due, at least in part, to deficient hepatic AFB1-detoxifying alpha-class glutathione S-transferases (GSTAs). Conversely, wild turkeys (Eastern wild, EW) are relatively resistant to the hepatotoxic, hepatocarcinogenic and immunosuppressive effects of AFB1 owing to functional gene expression and presence of functional hepatic GSTAs. This study was designed to compare the responses in gene expression in the gastrointestinal tract between DT (susceptible phenotype) and EW (resistant phenotype) following dietary AFB1 challenge (320 ppb for 14 days), specifically in cecal tonsil which functions in both nutrient absorption and gut immunity. RNAseq and gene expression analysis revealed significant differential gene expression in AFB1-treated animals compared to control-fed domestic and wild birds and in within-treatment comparisons between bird types. Significantly upregulated expression of the primary hepatic AFB1-activating P450 (CYP1A5) as well as transcriptional changes in tight junction proteins were observed in AFB1-treated birds. Numerous pro-inflammatory cytokines, TGF-&beta, and EGF were significantly down regulated by AFB1 treatment in DT birds and pathway analysis suggested suppression of enteroendocrine cells. Conversely, AFB1 treatment modified significantly fewer unique genes in EW birds, among these were genes involved in lipid synthesis and metabolism and immune response. This is the first investigation of the effects of AFB1 on the turkey gastro-intestinal tract. Results suggest that in addition to the hepatic transcriptome, animal resistance to this mycotoxin occurs in organ systems outside the liver, specifically as a refractory gastrointestinal tract.

Rumen protected Methionine (MET) and Lysine (LYS) are critical for milk protein synthesis in dairy cows. N-acetyl-L-methionine (NALM) is a MET derivative that consists of L-Met protected with an acetyl group that is attached to the α-amino group.N-acetyl-L-lysine (NALL) is a LYS derivative that is similarly protected. The objectives of these studies were to quantify the gastrointestinal availability of NALM and NALL. Three experiments were run as 3 × 3 Latin square using 3 second lactation Holstein cows that have been fitted with cannulas in the rumen and duodenum. The cows were fed diets containing the supplements for two weeks prior to each experiment so that the rumen microbes had time to adjust to the supplement. Each period consisted of 10 d of adaptation followed by 2 d of sampling. A dose of 0, 30, or 60 g of NALM was placed under the rumen mat at the time of feeding every day during experiment 1. The cows were similarly supplied with 0, 60, or 120 g of ƐNALL during experiment 2. The cows were supplemented with 0 g, 120 g ofƐNALL, or 120 g of diNALL during experiment 3. On day one of sampling, a liquid marker (Co-EDTA) was also administered at the time of the protected AA administration. Blood, ruminal, and duodenal samples were taken at hours 0, 1, 3, 6, 9, 12, and 24 post-feeding. There were no differences for milk production, milk protein, milk fat, or DMI for NALM or either NALL. There were no differences for ruminal escape (69.1% and 46.2% respectively) and duodenal appearance (2.16% and 3.40% respectively). The ruminal escape of ƐNALL was not different between the 120 g dose (32.7%) and the 60 g dose (27.2%). Duodenal appearance was higher (P < 0.01) for the 60 g dose (2.86%) than for the 120 g dose (1.19%) of ƐNALL. Acetate, propionate, butyrate, and valerate were higher (P < 0.01) for the supplemented cows during experiment 1 with NALL. There were no differences between ƐNALL and diNALL for rumen escape, duodenal appearance, VFA production, or blood LYS AUC. Results of the experiment verify significant protection of the N-acetyl MET and LYS from rumen degradation.

Osteoarthritis is a degenerative joint disease which plagues many older animals and humans. It is a disease that is characterized by the degeneration of joint cartilage, inflammation, as well as chronic pain and stiffness that results from this disorder. Unfortunately, most of the treatments for this painful and chronic disease mostly involve pain management and temporary relief strategies. These strategies usually include pain medication, non-steroidal anti-inflammatory drugs, injectable lubricants, as well as surgical techniques. However, because these treatments are just temporary fixes meant for pain management and to improve quality of life they must continue throughout the remainder of the animal's life. The ultimate goal of this, review is to compile knowledge about osteoarthritis. This will include everything from the clinical features and prevalence of the disease, as well as some of the possible causes of this disorder. Some of the most common treatments out there to treat pain and relieve inflammation, as well as some of the more experimental treatments that might not be approved for use yet, however they do still show some promise.

Editorial In the next two decades, the world population will increase significantly; the majority in the developing countries located in the tropics of Africa, Asia, Latin America, and the Caribbean. To feed such a population, it is necessary to increase the availability of food, particularly high-value animal protein foods produced locally, namely meat and dairy products. Dairy production in tropical regions has a lot of growth potential, but also poses a series of problems, particularly as dairy production systems were developed in temperate countries and in most cases are difficult to implement in the tropics. Drawbacks include hot weather and heat stress, the lack of availability of adequate feeds, poor infrastructure, and cold chain and the competition with cheap imports from temperate countries. This position paper reviews the major drawbacks in dairy production for the five major dairy species: cattle, water buffalo, sheep, goat, and camel, as well as the future trends in research and development. It also concerns the major trends in reproduction and production systems and health issues as well as environmental concerns, particularly those related to greenhouse gas emissions. Tropical Animal Health and Production now launches a topical collection on Tropical Dairy Science. We aim to publish interesting and significant papers in tropical dairy science. On behalf of the editorial board of the Tropical Animal Health and Production, we would like to invite all authors working in this field to submit their works on this topic to this topical collection in our journal info:eu-repo/semantics/publishedVersion

Menopause is associated with a decline in overall health in women. One health aspect impacted is glucose metabolism. As women experience menopause, their metabolism declines dramatically. The current study addressed the influence of ovarian somatic cells on the improvement of metabolic health through transplantations of young, germ cell-depleted ovaries. The purpose of this study is to expand the understanding of female reproductive health on metabolism. Control mice were grouped by age and treatment mice were age-matched. Treatment mice were placed into one of three groups: 1) mice received germ cell-depleted ovaries, 2) mice received germ cell-containing ovaries, and 3) mice received ovarian somatic cells via injection directly to their original ovary. All mice were subject to a glucose tolerance test, during which a bolus of dextrose was administered, and blood glucose levels were collected and recorded. Mice were euthanized between 680 and 700 days. Metabolic results showed an improvement of glucose metabolism in both germ cell-depleted and germ cell-containing groups compared to controls. No significance difference was noted between the germ cell-containing and germ cell-depleted groups. Somatic cell injection groups also showed improved glucose metabolism compared to controls. This experiment has shown that post-reproductive health is positively influenced by reproductive status. Additionally, somatic cells play an important role in the restoration of health to post-reproductive mice.

Fear-Free veterinary techniques are a new approach to caring for patients and clients in the veterinary clinic that have gained a great deal of popularity and traction. However, many veterinary professionals are left to question whether the utilization of these practices is resulting in a big enough change in client and patient stress levels to make it worth implementing these methods. This research looks into the views of veterinarians and clients on the effectiveness of these practices, and the competitive advantage of having a fear-free certified clinic from a business point of view.

Automatic Milking Systems (AMS) – or robotic milkers – have been on the market for over twenty years. However, AMS introduction is relatively new to the Western U.S., and requires challenging adjustments and new management systems., https://digitalcommons.usu.edu/fsrs2018/1005/thumbnail.jpg

Arenaviruses are a significant cause of hemorrhagic fever, an often-fatal disease for which there is no approved antiviral therapy. Lassa fever in particular generates high morbidity and mortality in West Africa, where the disease is endemic, and a recent outbreak in Nigeria was larger and more geographically diverse than usual. We are developing LHF-535, a small-molecule viral entry inhibitor that targets the arenavirus envelope glycoprotein, as a therapeutic candidate for Lassa fever and other hemorrhagic fevers of arenavirus origin. Using a lentiviral pseudotype infectivity assay, we determined that LHF-535 had sub-nanomolar potency against the viral envelope glycoproteins from all Lassa virus lineages, with the exception of the glycoprotein from the LP strain from lineage I, which was 100-fold less sensitive than that of other strains. This reduced sensitivity was mediated by a unique amino acid substitution, V434I, in the transmembrane domain of the envelope glycoprotein GP2 subunit. This position corresponds to the attenuation determinant of Candid#1, a live-attenuated Junín virus vaccine strain used to prevent Argentine hemorrhagic fever. Using a virus-yield reduction assay, we determined that LHF-535 potently inhibited Junín virus, but not Candid#1, and the Candid#1 attenuation determinant, F427I, regulated this difference in sensitivity. We also demonstrated that a daily oral dose of LHF-535 at 10 mg/kg protected mice from a lethal dose of Tacaribe virus. Serial passage of Tacaribe virus in LHF-535-treated Vero cells yielded viruses that were resistant to LHF-535, and the majority of drug-resistant viruses exhibited attenuated pathogenesis. These findings provide a framework for the clinical development of LHF-535 as a broad-spectrum inhibitor of arenavirus entry and provide an important context for monitoring the emergence of drug-resistant viruses., Author summary Lassa fever is a viral hemorrhagic fever disease that is transmitted to humans primarily through contact with the urine or feces of infected rodents. The disease is endemic in West Africa, and an unusually large outbreak occurred in Nigeria in early 2018. The case fatality rate was 25% among confirmed cases, underscoring the need for an effective antiviral therapy. Here, we evaluated the small-molecule drug LHF-535, which targets the arenavirus envelope glycoprotein, for broad-spectrum activity against Lassa viruses of different lineages and related arenaviruses that cause hemorrhagic fever diseases in South America. We also selected for LHF-535-resistant viruses and characterized their genotype and phenotype. Using a combination of surrogate systems and wild-type viruses, we determined that all tested Lassa virus strains and New World hemorrhagic fever arenaviruses were sensitive to LHF-535. Sensitivity to the drug was modulated by specific amino acid changes in the viral envelope glycoprotein, and the majority of emerging drug-resistant viruses were attenuated for virulence. Similarly, the live-attenuated vaccine strain for Argentine hemorrhagic fever was also resistant to LHF-535. These findings indicate that LHF-535 targets a viral virulence determinant, the mutation of which may result in the emergence of drug-resistant viruses, but with reduced capacity for virulence.

Although a highly effective vaccine is available, the number of yellow fever cases has increased over the past 2 decades, which highlights the pressing need for antiviral therapeutics. In a high-throughput screening campaign, we identified an acetic acid benzodiazepine (BDAA) compound which potently inhibits yellow fever virus (YFV). Interestingly, while treatment of YFV-infected cultures with 2 μM BDAA reduced the virion production by greater than 2 logs, the compound was not active against 21 other viruses from 14 different viral families. Selection and genetic analysis of drug-resistant viruses revealed that replacement of the proline at amino acid 219 (P219) of the nonstructural protein 4B (NS4B) with serine, threonine, or alanine conferred YFV with resistance to BDAA without apparent loss of replication fitness in cultured mammalian cells. However, replacement of P219 with glycine conferred BDAA resistance with significant loss of replication ability. Bioinformatics analysis predicts that the P219 amino acid is localized at the endoplasmic reticulum lumen side of the fifth putative transmembrane domain of NS4B, and the mutation may render the viral protein incapable of interacting with BDAA. Our studies thus revealed an important role and the structural basis for the NS4B protein in supporting YFV replication. Moreover, in YFV-infected hamsters, oral administration of BDAA protected 90% of the animals from death, significantly reduced viral load by greater than 2 logs, and attenuated virus infection-induced liver injury and body weight loss. The encouraging preclinical results thus warrant further development of BDAA or its derivatives as antiviral agents to treat yellow fever. IMPORTANCE Yellow fever is an acute viral hemorrhagic disease which threatens approximately 1 billion people living in tropical areas of Africa and Latin America. Although a highly effective yellow fever vaccine has been available for more than 7 decades, the low vaccination rate fails to prevent outbreaks in at-risk regions. It has been estimated that up to 1.7 million YFV infections occur in Africa each year, resulting in 29,000 to 60,000 deaths. Thus far, there is no specific antiviral treatment for yellow fever. To cope with this medical challenge, we identified a benzodiazepine compound that selectively inhibits YFV by targeting the viral NS4B protein. To our knowledge, this is the first report demonstrating in vivo safety and antiviral efficacy of a YFV NS4B inhibitor in an animal model. We have thus reached a critical milestone toward the development of specific antiviral therapeutics for clinical management of yellow fever.

Placenta-derived extracellular vesicles (EVs) are involved in communication between the placenta and maternal immune cells possibly leading to a modulation of maternal T-cell signaling components. The ability to identify EVs in maternal blood may lead to the development of diagnostic and treatment tools for pregnancy complications. The objective of this work was to differentiate EVs from bovine placenta (trophoblast) and peripheral blood mononuclear cells (PBMC) by a label-free, non-invasive Raman spectroscopy technique. Extracellular vesicles were isolated by ultracentrifugation. Dynamic light scattering (DLS) and scanning electron microscopy (SEM) were applied to verify the presence and the size distribution of EVs. Raman peaks at 728 cm-1 (collagen) and 1573 cm-1 (protein) were observed only in PBMC-derived EVs, while the peaks 702 cm-1 (cholesterol) and 1553 cm-1 (amide) appeared only in trophoblast-derived EVs. The discrimination of the Raman spectral fingerprints for both types of EVs from different animals was performed by principal component analysis (PCA) and linear discriminant analysis (LDA). The PCA and LDA results clearly segregated the spectral clusters between the two types of EVs. Moreover, the PBMC-derived EVs from different animals were indistinguishable, while the trophoblast-derived EVs from three placental samples of different gestational ages showed separate clusters. This study reports for the first time the Raman characteristic peaks for identification of PBMC and trophoblast-derived EVs. The development of this method also provides a potential tool for further studies investigating the causes and potential treatments for pregnancy complications.

Consumption of a Western type diet is a known risk factor for colorectal cancer. Our group previously developed the total Western diet (TWD) for rodents with energy and nutrient profiles that emulate a typical Western diet. In this study, we tested the hypothesis that consumption of the TWD would enhance colitis, delay recovery from gut injury and promote colon tumorigenesis. In multiple experiments using the azoxymethane + dextran sodium sulfate or ApcMin/+ mouse models of colitis-associated colorectal carcinogenesis (CAC), we determined that mice fed TWD experienced more severe and more prolonged colitis compared to their counterparts fed the standard AIN93G diet, ultimately leading to markedly enhanced colon tumorigenesis. Additionally, this increased tumor response was attributed to the micronutrient fraction of the TWD, and restoration of calcium and vitamin D to standard amounts ameliorated the tumor-promoting effects of TWD. Finally, exposure to the TWD elicited large scale, dynamic changes in mRNA signatures of colon mucosa associated with interferon (IFN) response, inflammation, innate immunity, adaptive immunity, and antigen processing pathways, among others. Taken together, these observations indicate that consumption of the TWD markedly enhanced colitis, delayed recovery from gut injury, and enhanced colon tumorigenesis likely via extensive changes in expression of immune-related genes in the colon mucosa.

Clinical evidence suggests that Zika virus contributes to Guillain-Barré syndrome that causes temporary paralysis. We utilized a recently described Zika virus mouse model of temporary flaccid paralysis to address the hypothesis that treatment with an N-methyl-D-aspartate receptor antagonist, memantine, can reduce the incidence of paralysis. Aged interferon alpha/beta-receptor knockout mice were used because of their sublethal susceptibility to Zika virus infection. Fifteen to twenty-five percent of mice infected with a Puerto Rico strain of Zika virus develop acute flaccid paralysis beginning at days 8–9 and peaked at days 10–12. Mice recover from paralysis within a week of onset. In two independent studies, twice daily oral administration of memantine at 60 mg/kg/day on days 4 through 9 after viral challenge significantly reduced the incidence of paralysis. No efficacy was observed with treatments from days 9 through 12. Memantine treatment in cell culture or mice did not affect viral titers. These data indicate that early treatment of memantine before onset of paralysis is efficacious, but treatments beyond the onset of paralysis were not efficacious. The effect of this N-methyl-D-aspartate receptor antagonist on the incidence of Zika virus-induced paralysis may provide guidance for investigations on the mechanism of paralysis.

Zika virus (ZIKV) is an arboviral infection that may be sexually transmitted. The present study aims to determine if accessory sex glands are a potential source of infectious virus and important in sexual transmission. Male interferon type I receptor knockout (Ifnar−/−) mice were challenged subcutaneously with a Puerto Rican ZIKV isolate. Reproductive tissues were harvested seven days after viral challenge and artificial insemination fluid derived from epididymis or homogenized accessory sex glands (seminal plasma) was obtained. Naïve interferon type I and II receptor knockout (AG129) females were pre-treated with progesterone, and inoculated intravaginally with either epididymal flush or seminal plasma from ZIKV-infected males. ZIKV RNA was demonstrated in the artificial insemination fluid and ZIKV antigen was detected in epididymal epithelial cells but not within seminiferous tubules at the time of artificial insemination fluid collection. Peripheral viremia, demonstrated by ZIKV RNA in whole blood samples of females from each challenge group was observed. Infectious virus was present in both epididymal fluid and seminal plasma. These studies provide evidence of passage of virus from epididymal flush and seminal plasma to naïve females via artificial insemination and provides a model for the study of sexual transmission of ZIKV.

Mastitis in a single quarter can cause high somatic cell counts (SCC), clinical mastitis, and death in dairy cows. Currently, management of these mastitic quarters presents a problem for the dairy industry. Casein hydrolysate (CH) is an intramammary (IMM) infusion treatment reported to induce mammary involution. The primary objectives of this study were to investigate whether IMM CH treatment of single high SCC quarters, followed by cessation of quarter milk production for the remainder of lactation, was effective in reducing cow–level SCC and whether that quarter resumed milk production following calving. Three treatment groups were used: CH, non-hydrolyzed casein (NHC), and cessation of milking only (negative; N). Treatments were assigned in a 2:2:1 ratio for 40 cows enrolled in the study; 27 cows completed the entire protocol. Following IMM infusion and involution of the single mastitic quarter, decreases in cow–level SCC (-966,000/ml) and milk production (-11 lb (5 kg), -14%) with 3 remaining lactating quarters were significant for all 28 cows combined. Cows treated with CH (n=17) had a significant decrease in cow– level SCC (-1,150,000/ml) during remaining lactation. All treated quarters returned to milk production after calving, and their proportion of total–cow milk production (24%) was not different than before treatment (28%). After calving, treated quarters’ decrease in SCC was significant for CH (-2,763,000/ml; n=14) and N (–5,324,000/ml; n=5). Of 16 quarters with positive milk culture before treatment that completed the protocol, 88% (14/16) were cured (no isolation of the same bacteria for 3 weeks following calving). A new intramammary infection (IMI) was detected in 67% (18/27) of previously treated quarters post-calving. Infusing single mastitic quarters with casein hydrolysate to induce involution for the remainder of lactation may be a promising alternative to current methods.

The highly efficacious live-attenuated 17D yellow fever (YF) vaccine is occasionally associated with rare life-threatening adverse events. Modified Vaccinia Ankara (MVA), a non-replicating poxvirus, has been used as a vaccine platform to safely deliver various antigens. A MVA-based YF vaccine (MVA-BN-YF) was tested with and without a non-mineral oil adjuvant in a hamster model of lethal YF disease and protective efficacy of this vaccine was compared with the 17D vaccine. The vaccine candidate MVA-BN-YF generated a protective response in hamsters infected with YFV that was comparable to protection by the live 17D vaccine. Similar levels of neutralizing antibody were observed in animals vaccinated with either vaccine alone or vaccine with adjuvant. Significant improvement in survival, weight change, and serum alanine aminotransferase (ALT) levels were observed in vaccinated hamsters when administered 42 and 14 days prior to challenge with Jimenez YF virus (YFV). Neutralizing antibodies induced by MVA-BN-YF were transferred to naive hamsters prior to virus challenge. Passive administration of neutralizing antibody 24 hours prior to virus infection resulted in significantly improved survival and weight change. A trend towards reduced liver enzyme levels was also observed. MVA-BN-YF, therefore, represents a safe alternative to vaccination with live attenuated YF virus.

The mammary gland is a very important organ for reproduction in mammals because it produces milk which serves as the primary source of nutrients for newly-born offspring. Previous studies suggest that its development is regulated by circadian clocks, biochemical oscillators that generate circadian rhythms (the body’s internal clock). The circadian system plays a major role in homeostasis, coordinating the body’s internal physiology and synchronizing it with the external environment. Our lab showed that levels of the BMAL1 protein, a core clock component, increased in the mammary gland at the onset of lactation. Treatment of mammary epithelial cells (HC11) with the hormone prolactin significantly increased BMAL1 levels. We hypothesize that the secretion of prolactin during lactogenesis induces expression of BMAL1 in the mouse mammary gland through the STAT5a signaling pathway. The objective of the project was to determine the effect of different amounts of STAT5a protein on BMAL1 levels with and without prolactin treatment. For this experiment, western blot analysis was used to measure STAT5a and BMAL1 levels in wild type HC11 cells and in HC11 cell lines that were genetically modified to: 1) express very high levels of STAT5a (STAT5a-OE), 2) express a mutant form of STAT5a that is inactive (STAT5a-dnl), and 3) delete the BMAL1 gene (BMAL1 KO). Our first round of analysis showed that overexpressing STAT5a increased BMAL1 protein levels, especially in cells differentiated by prolactin. Results from this experiment would allow us to better understand the relationship between mammary gland development and the circadian system.

Lactose in milk has relatively low variation regardless of season, breed, or country. The study of lactose concentration and correlation among other milk components is limited. Furthermore, dairy farmers have limited access to the lactose data and are not familiar with it. This study was conducted to: 1) investigate the phenotypic correlation between lactose and other milk components; and 2) determine the importance of lactose for dairy herds. Monthly DHIA records from Utah (DHIA), Dairy Herd Performance Test (DHTP) records from Ibaraki, Japan, and California herd average data (CHAD) covering 27 states were used to analyze the relationships between milk lactose concentrations and parameters related to milk production. Record spans for each data sets for DHIA, DHTP and CHAD were 7 consecutive years ending August 2017, 12 consecutive years ending August 2017, and 9 consecutive years ending August 2017 respectively. Means for the DHIA records were 187.9 days in milk, 34.6 kg/d milk yield, 4.08% lactose, 3.88% fat, 3.15% protein, 8.86% SNF, 14.4 mg/dl MUN, 256,000 SCC, and 2.4 parity. A negative correlation was observed between lactose concentration and parity (r = -0.27). Positive phenotypic correlations between lactose percentage and milk yield (r = 0.28), and negative correlations with fat (r = -0.17), protein (r = -0.21) and SCC (r = -0.30) were observed. No seasonal fluctuations of milk lactose concentrations were observed. Mean for the DHTP records were 4.55% lactose, 3.95% fat, 3.29% protein, 8.74% SNF, 283 mOsm/kg osmotic pressure, 19,600 bacteria count, and 222,000 SCC. A seasonality in the lactose concentrations were observed, with the lowest concentrations of lactose observed in late summer and fall. There were no phenotypic correlations between lactose concentrations and protein, but a negative correlation with fat (r = -0.12) was observed. A significant negative correlation with SCC (r = -0.28) was also observed. Based on the CHAD records, cows with lower lactose concentrations showed a higher culling rate, which indicates that the milk lactose concentration could be a potential indicator of problems in a dairy herd. This study suggest that lactose level could be an indicator for managing dairy farms. Dairy farmers should be able to access lactose data, and these data should be provided by DHIA.

The non-classical MHC class I (MHC-Ib) proteins are important modulators of immune system during pregnancy favoring survival of the fetus. Contrary to ubiquitously expressed classical MHC-I proteins, MHC-Ib proteins are oligomorphic, and expressed in specific cell/tissue types thus minimizing maternal immune-mediated rejection of fetal-allograft and a successful pregnancy. A unique characteristic of MHC-Ib glycoproteins is expression of surface and soluble isoforms due to alternative splicing phenomenon. Bovine fetal trophoblast cells, during the third trimester of pregnancy, express non-classical bovine leukocyte antigen class Ib (BoLA-Ib) antigens. BoLA-NC3*00101, is a non-classical class I allele from cattle AH11 haplotype and is expressed as cell surface isoform. However, lack of monoclonal antibody (mAb) hinders the development of specific assay to detect the soluble/secreted BoLA-I antigens released by fetal trophoblast cells. The objective of this study is to synthesize mAb specific to NC3*00101 molecule to develop an isotype-matched ELISA to assess BoLA-I protein(s). We demonstrated that majority of NC3*00101 hybridoma-supernatants were low-titer and showed inappreciable reactivity in ELISA and flow cytometry assays. We identified a clone of hybridoma (NC3-3*2) that secreted mAb specific to BoLA-NC3*00101 as demonstrated with flow cytometry. NC3-3*2 clone secreted supernatant that captured BoLA-NC3*00101 protein in ELISA. Unfortunately, despite several efforts we could not recover the cryopreserved hybridoma. Non-recoverability and instability, thus, limited production of NC3*00101-mAbs. This study provides the evidence that mice immunized with bovine class Ib proteins elicit a specific immunogenic response and warrants future studies into generation of stable hybridoma secreting antibodies against BoLA-Ib proteins using robust methods of fusion and cryopreservation of hybridomas.

Japanese encephalitis virus (JEV), a mosquito-borne zoonotic flavivirus, is an enveloped positive-strand RNA virus that can cause a spectrum of clinical manifestations, ranging from mild febrile illness to severe neuroinvasive disease. Today, several killed and live vaccines are available in different parts of the globe for use in humans to prevent JEV-induced diseases, yet no antivirals are available to treat JEV-associated diseases. Despite the progress made in vaccine research and development, JEV is still a major public health problem in southern, eastern, and southeastern Asia, as well as northern Oceania, with the potential to become an emerging global pathogen. In viral replication, the entry of JEV into the cell is the first step in a cascade of complex interactions between the virus and target cells that is required for the initiation, dissemination, and maintenance of infection. Because this step determines cell/tissue tropism and pathogenesis, it is a promising target for antiviral therapy. JEV entry is mediated by the viral glycoprotein E, which binds virions to the cell surface (attachment), delivers them to endosomes (endocytosis), and catalyzes the fusion between the viral and endosomal membranes (membrane fusion), followed by the release of the viral genome into the cytoplasm (uncoating). In this multistep process, a collection of host factors are involved. In this review, we summarize the current knowledge on the viral and cellular components involved in JEV entry into host cells, with an emphasis on the initial virus-host cell interactions on the cell surface.

Legumes that contain condensed tannins may have lower ruminal protein degradation than alfalfa. The present study investigated the effects of feeding birdsfoot trefoil (Lotus corniculatus L.) hay on lactational performance and N utilization and excretion. Eight multiparous Holstein cows in midlactation (150 ± 22.3 d-in-milk) were randomly assigned to 2 treatments [alfalfa hay-based total mixed ration (AHT) or birdsfoot trefoil hay-based total mixed ration (BHT)] in a crossover design with 2 experimental periods. Each experimental period lasted 17 d (14 d of adaptation and 3 d of sampling and total collection). Hays comprised approximately 50% of DM in experimental diets. There were no treatment effects on dry matter intake (DMI; 21.4 vs. 20.7 kg/d), milk yield (29.4 vs. 28.1 kg/d), milk fat concentration (3.20% vs. 3.21%), and milk protein concentration (3.20% vs. 3.16%) for AHT and BHT, respectively. In addition, dietary treatments did not affect milk yield/DMI or energy-corrected milk yield/DMI. In contrast, apparent crude protein digestion decreased in cows fed BHT compared with those fed AHT (60.7% vs. 69.1%). Concentration of milk urea-N decreased by feeding BHT compared with AHT (11.9 vs. 13.3 mg/100 mL), whereas total N excretion did not differ between AHT and BHT diets. However, cows fed BHT excreted more N in feces (194 vs. 168 g/d), whereas urinary N excretion was lower compared with cows fed AHT. The shift of N to feces resulted in a decrease in urinary N:fecal N ratio in cows fed BHT relative to those fed AHT. Overall results in the current study suggest that feeding birdsfoot trefoil in dairy diets shifts routes of N from urine to feces compared with feeding alfalfa hay, with little effect on lactational performance. Reduction in urinary N and any impact on environment may be attributed to functional effect of condensed tannins in birdsfoot trefoil hay.

It is our great pleasure to welcome you to the second Large Animal Genetic Engineering (LAGE) Summit in beautiful Park City, Utah. We hope that you enjoy the meeting and take advantage of your free time to get out and see what Park City has to offer. Our objectives for the summit are twofold: (1) to provide updates on genetic engineering technologies, including genome editing, and their biomedical and agricultural applications in academia and industry; and (2) to engage federal agencies, industry representatives and academic researchers in a policy development forum. We hope that this year’s meeting will build on the success of the first LAGE Summit, which was held in Bethesda, Maryland, in 2016. We are grateful to all of our speakers and panel participants who have agreed to participate in the meeting. Most importantly, we thank our sponsors who have helped make the summit possible and the people who have worked hard to make sure that the summit is a success. We trust that the summit will be informative, invigorating and intellectually stimulating. We also hope that this event will provide an opportunity for you to renew old friendships, as well as develop new contacts and collaborations.

The accumulating evidence demonstrates that Syrian hamsters have advantages as models for various diseases. To develop a Syrian hamster (Mesocricetus auratus) model of human immunodeficiency caused by RAG1 gene mutations, we employed the CRISPR/Cas9 system and introduced an 86-nucleotide frameshift deletion in the hamster RAG1 gene encoding part of the N-terminal non-core domain of RAG1. Histological and immunohistochemical analyses demonstrated that these hamsters (referred herein as RAG1-86nt hamsters) had atrophic spleen and thymus, and developed significantly less white pulp and were almost completely devoid of splenic lymphoid follicles. The RAG1-nt86 hamsters had barely detectable CD3+ and CD4+ T cells. The expression of B and T lymphocyte-specific genes (CD3γ and CD4 for T cell-specific) and (CD22 and FCMR for B cell-specific) was dramatically reduced, whereas the expression of macrophage-specific (CD68) and natural killer (NK) cell-specific (CD94 and KLRG1) marker genes was increased in the spleen of RAG1-nt86 hamsters compared to wildtype hamsters. Interestingly, despite the impaired development of B and T lymphocytes, the RAG1-86nt hamsters still developed neutralizing antibodies against human adenovirus type C6 (HAdV-C6) upon intranasal infection and were capable of clearing the infectious viruses, albeit with slower kinetics. Therefore, the RAG1-86nt hamster reported herein (similar to the hypomorphic RAG1 mutations in humans that cause Omenn syndrome), may provide a useful model for studying the pathogenesis of the specific RAG1-mutation-induced human immunodeficiency, the host immune response to adenovirus infection and other pathogens as well as for evaluation of cell and gene therapies for treatment of this subset of RAG1 mutation patients.

Bovine mastitis, or inflammation of the mammary gland, has substantial economic and animal welfare implications. A genetic basis for mastitis resistance traits is recognized and can be used to guide selective breeding programs. The discovery of regions of the genome associated with mastitis resistance, and knowledge of the underlying molecular mechanisms responsible, can facilitate development of efficient mastitis control and therapeutic strategies. The objectives of this dissertation research were to identify sites of genetic variation associated with mastitis resistance, and to define the contributions of the milk-secreting epithelial cells to mammary gland immune responses and mastitis resistance. Twenty seven regions of the bovine genome potentially involved in mastitis resistance were identified in Holstein dairy cattle. Additionally, this research demonstrates a role of bovine mammary epithelial cells in mastitis resistance, and provides guidance for the use of an in vitro model for mastitis studies. Primary bovine mammary epithelial cells from mastitis-resistant cows have differential expression of 42 inflammatory genes compared with cells from mastitis-susceptible cows, highlighting the importance of epithelial cells in mastitis resistance. Bovine mammary epithelial cells display both similarities and differences in pro-inflammatory gene expression compared to fibroblasts, and their expression of inflammatory genes is influenced by administration of the enzyme phospholipase A2. The growth potential of milk-derived bovine mammary epithelial cells in vitro can be extended, facilitating their use in mastitis studies, by transfection with a viral protein. Collectively, this research contributes to current knowledge on bovine mastitis resistance and in vitro models.