Animal & Avian Sciences
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Item Variations in Vocalizations of Fin Whales, Balaenoptera physalus, in the St. Lawrence River(1980) Edds, Peggy Louise; Buchler, Edward; Animal & Avian Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, MD)Recordings were made of vocalizations from fin whales, Balaenoptera physalus from a fixed hydrophone in the St. Lawrence River between 28 June and 27 September 1979. Land-based observers monitored activity from a hillside hut while recordings were being made from a shoreline site. Photographs of dorsal fin variations were used to identify distinctive individuals. The 1522 sounds recorded were classified into 11 categories based on frequency and temporal characteristics. The predominant call was a descending sweep of frequencies. Parameters measured for this downsweep exhibited a dichotomy of characteristics which indicates calls with initial frequencies below 40 Hz have less variability than calls with initial frequencies above 40 Hz. In general, solitary animals produced primarily the lower frequency downsweeps. Higher frequency downsweeps were recorded from pairs or trios of fin whales. Solitary individuals did not exhibit unique variations in downsweep parameters. No clusters of values which might indicate uniquely individual ranges were consistently present in multiple animal recordings. The data suggest that the variability of fin whale vocalizations is primarily contextual rather than individual.Item Infant and Juvenile-Directed Care Behaviors in Adult Toque Macaques, Macaca Sinica(1985) Baker-Dittus, Anne; Animal & Avian Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, MD)The identity of care-givers, and the distribution of care to young are examined in a free-ranging polygynous primate, Macaca sinica. Care behavior is used as one measure of investment in young. There is no evidence that dominance rank influences offspring sex, but high-ranking mothers provide more care to newborn infants than do low-ranking mothers. Mothers provide more care to infant and first year sons than to daughters. This supports Dittus's suggestion (1979, 1980) that mothers should schedule care to sons and daughters differently, providing high levels of care to sons before they emigrate from the natal group. Care to daughters is low per unit time, but continues over a long period because daughters remain in the natal group. Adult females, other than the mother, tolerate, groom and support young. These behaviors are low cost, relative to the high cost behaviors of nursing and carrying, which only mothers provide. Young tend to associate with adult female relatives. Unlike male-biased maternal care, adult female care is biased towards female young; and female young return care more than do male young . Adult female rank is positively correlated with the amount of support adult females provide to female young, and the amount of grooming they receive from female young. Adult female rank has no effect on any measure of association between adult females and male young. Taken together these findings suggest that reciprocal altruism has been important in shaping adult female interactions with young. Adult and subadult M. sinica males direct the low cost, affiliative behaviors of hugging, carrying and grooming towards male infants; and direct aggressive behaviors towards female infants. I suggest that the distribution of behaviors reflects their value to male and female young. Affiliative behaviors to male infants, and aggressive behaviors to female infants serve to channel infants into association with adults from whom they can learn appropriate sex-typical behaviors, and as such benefit both male and female infants. High cost care behaviors of support in agonistic interactions, and protection are provided only by the adult male, and are provided to male and female young equally.Item The interaction between a parasitic barnacle, Loxothylacus panopaei (Cirripedia, Rhizocephala), and three of its crab host species (Brachyura, Xanthidae) along the east coast of North America.(1993) Alvarez, Fernando; Reaka-Kudla, Marjorie; Animal & Avian Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, MD)Rhizocephalan barnacles parasitize primarily decapod crustaceans (Bocquet-Vedrine 1968, Overstreet 1983). The principal effect of the parasites is often the permanent sterilization of the hosts (Bocquet-Vedrine 1972, O'Brien & Van Wyk 1984). Regardless of the processes involved, the end result of the infection is the creation of non-reproductive individuals that will use resources that otherwise would be available to the healthy traction of the host population. The potential impact on the host population then resides in the accumulation of sterile resource-using individuals in each generation. Prevalences of barnacles on anomuran and brachyuran crabs have been reported in a number of studies (e.g., Walker 1985, Hawkes et al. 1986, Johnson et al. 1986, Wardle & Tirpak 1991 ). Rhizocephalan infections can reach very high levels locally, suggesting that entire host populations may have a greatly reduced reproductive capacity. However, in most host species, the pattern of distribution of parasites is not uniform throughout the host's range. I examined the crab-rhizocephalan interaction using the system composed by the barnacle Loxothylacus panopaei and three of its crab host species: Panopeus lacustris, Eurypanopeus depressus, and Rhithropanopeus harrisii, occurring along the east coast of the North America. Through experimental infections conducted in the laboratory, it was determined that B. harrisii of a wide range of sizes could be infected by L. panopaei. The complete life-cycle of the parasite was manipulated in the laboratory. Neither host molting frequency nor host molt increments differed significantly between parasitized and control crabs. Host survival was significantly reduced during the parasite's developmental period; the heaviest mortality of the host occurred in the megalopal stage.Item MOLECULAR ANALYSIS OF CYCLOPHILIN FUNCTION IN THE YEAST SACCHAROMYCES CEREVISIAE(1994) Davis, Edward S.; Brennan, Miles S.; Digital Repository at the University of Maryland; University of Maryland (College Park, MD)The cyclophilins are a family of proteins first identified as receptors for cyclosporin A (CsA), a cyclic peptide of fungal origin. CsA inhibits T-lymphocyte activation, and is thus a potent immunosuppressant. Although cyclophilins are ubiquitous, and highly conserved, among eukaryotes, their normal physiological functions are unknown. As the receptors for CsA, cyclophilins might be involved in regulating signal transduction pathways. Cyclophilns also have peptidyl-prolyl, cis-trans isomerase (PPIase) activity in vitro, suggesting a role in protein folding in vivo. While CsA inhibits cyclophilin's PPIase activity, this inhibition is insufficient to account for the pharmacological activity of CsA. Therefore, previous results cannot be readily synthesized into a model for cyclophilin function. The goal of this project was to define and characterize physiological roles of cyclophilins using the yeast S. cerevisiae. Three S. cerevisiae cyclophilin genes were cloned and inactivated by insertional mutagenesis. I demonstrated that one, CPR3, is necessary for the efficient metabolism of non-fermentable carbon sources. The CPR] gene product, Cpr3, is localized to the mitochondrial matrix, and a truncated version of Cpr3 expressed in bacteria binds CsA. CPR3 inactivation does not significantly compromise the induction of transcription of two nuclear cytochrome genes. Thus, Cpr3 is not necessary for the signal transduction pathway governing cytochrome gene expression. To identify biochemical targets of Cpr3, I demonstrated that inactivation of a mitochondrial lactate dehydrogenase is insufficient to account for the growth defect of cpr3 mutants. An exhaustive search for high-copy suppressors of the growth defect of cpr3 mutants led to the identification of a novel gene, JEN1, that suppresses the growth defect at elevated temperature. JEN1 encodes a protein that is probably a lactate transporter, and thus not a direct biochemical target of Cpr3. A dominant mutation in a nuclear gene, JEN2, suppresses the growth defect of cpr3 mutants on lactate at 30°C and 37°C. JEN2 might encode a direct biochemical target of Cpr3. In summary, the cyclophilin, Cpr3, plays a general role in the efficient function of yeast mitochondria, and presents an excellent model system for studying cyclophilin function.Item Targeted Gene Knockin in Porcine Somatic Cells Using CRISPR/Cas Ribonucleoproteins(MDPI, 2016-05-26) Park, Ki-Eun; Park, Chi-Hun; Powell, Anne; Martin, Jessica; Donovan, David M.; Telugu, Bhanu P.The pig is an ideal large animal model for genetic engineering applications. A relatively short gestation interval and large litter size makes the pig a conducive model for generating and propagating genetic modifications. The domestic pig also shares close similarity in anatomy, physiology, size, and life expectancy, making it an ideal animal for modeling human diseases. Often, however, the technical difficulties in generating desired genetic modifications such as targeted knockin of short stretches of sequences or transgenes have impeded progress in this field. In this study, we have investigated and compared the relative efficiency of CRISPR/Cas ribonucleoproteins in engineering targeted knockin of pseudo attP sites downstream of a ubiquitously expressed COL1A gene in porcine somatic cells and generated live fetuses by somatic cell nuclear transfer (SCNT). By leveraging these knockin pseudo attP sites, we have demonstrated subsequent phiC31 integrase mediated integration of green fluorescent protein (GFP) transgene into the site. This work for the first time created an optimized protocol for CRISPR/Cas mediated knockin in porcine somatic cells, while simultaneously creating a stable platform for future transgene integration and generating transgenic animals.Item Somatic Cell Nuclear Transfer Followed by CRIPSR/Cas9 Microinjection Results in Highly Efficient Genome Editing in Cloned Pigs(MDPI, 2016-12-03) Sheets, Timothy P.; Park, Chi-Hun; Park, Ki-Eun; Powell, Anne; Donovan, David M.; Telugu, Bhanu P.The domestic pig is an ideal “dual purpose” animal model for agricultural and biomedical research. With the availability of genome editing tools such as clustered regularly interspaced short palindromic repeat (CRISPR) and associated nuclease Cas9 (CRISPR/Cas9), it is now possible to perform site-specific alterations with relative ease, and will likely help realize the potential of this valuable model. In this article, we investigated for the first time a combination of somatic cell nuclear transfer (SCNT) and direct injection of CRISPR/Cas ribonucleoprotein complex targeting GRB10 into the reconstituted oocytes to generate GRB10 ablated Ossabaw fetuses. This strategy resulted in highly efficient (100%) generation of biallelic modifications in cloned fetuses. By combining SCNT with CRISPR/Cas9 microinjection, genome edited animals can now be produced without the need to manage a founder herd, while simultaneously eliminating the need for laborious in vitro culture and screening. Our approach utilizes standard cloning techniques while simultaneously performing genome editing in the cloned zygotes of a large animal model for agriculture and biomedical applications.Item Characterization of Copy Number Variation’s Potential Role in Marek’s Disease(MDPI, 2017-05-09) Xu, Lingyang; He, Yanghua; Ding, Yi; Sun, Guirong; Carrillo, Jose Adrian; Li, Yaokun; Ghaly, Mona M.; Ma, Li; Zhang, Huanmin; Liu, George E.; Song, JiuzhouMarek’s Disease (MD) is a highly contagious pathogenic and oncogenic disease primarily affecting chickens. Chicken Lines 63 and 72, as well as their recombinant congenic strains (RCS) with varied susceptibility to MD, are ideal models to study the complex mechanisms of genetic resistance to MD. In this study, we investigated copy number variation (CNV) in these inbred chicken lines using the Affymetrix Axiom HD 600 K SNP genotyping array. We detected 393 CNV segments across all ten chicken lines, of which 12 CNVs were specifically identified in Line 72. We then assessed genetic structure based on CNV and observed markedly different patterns. Finally, we validated two deletion events in Line 72 and correlated them with genes expression using qPCR and RNA-seq, respectively. Our combined results indicated that these two CNV deletions were likely to contribute to MD susceptibility.Item Antimicrobial Effect and Probiotic Potential of Phage Resistant Lactobacillus plantarum and its Interactions with Zoonotic Bacterial Pathogens(MDPI, 2019-06-05) Nagarajan, Vinod; Peng, Mengfei; Tabashsum, Zajeba; Salaheen, Serajus; Padilla, Joselyn; Biswas, DebabrataDevelopment of phage-resistant probiotic particularly Lactobacillus is an alternative approach to enhance their beneficial effects as in animal feed supplements. In this study, we developed phage-resistant Lactobacillus plantarum (LP+PR) mutant and compared their antimicrobial effects and probiotic potential against zoonotic bacterial pathogens including Salmonella enterica serovar Typhimurium, enterohemorrhagic Escherichia coli (EHEC), Staphylococcus aureus, and Listeria monocytogenes with phage-sensitive L. plantarum (LP) strain. LP+PR strain showed markedly higher growth rate than wild-type LP strain. In co-culture with LP+PR and in the presence of cell-free cultural supernatants (CFCSs) of LP+PR, the growth of S. Typhimurium, EHEC, S. aureus, and L. monocytogenes were reduced significantly (P < 0.05). The adhesion ability of LP+PR was slightly higher than the LP on human epithelial INT-407 cells. Most importantly, LP+PR strain significantly inhibited the adhesive and invasive abilities of all four zoonotic pathogens to INT-407 cells (P < 0.05). Moreover, real-time qPCR revealed that in the presence of LP+PR strain or its CFCSs, expression of virulence genes of these zoonotic bacterial pathogens were suppressed significantly (P < 0.05). These findings suggest that the LP+PR strain is capable of inhibiting major zoonotic bacterial pathogens efficiently and would be a potential candidate for industrial usage in animal production or fermentation.Item Marek’s Disease Virus Infection Induced Mitochondria Changes in Chickens(MDPI, 2019-06-27) Chu, Qin; Ding, Yi; Cai, Wentao; Liu, Lei; Zhang, Huanmin; Song, JiuzhouMitochondria are crucial cellular organelles in eukaryotes and participate in many cell processes including immune response, growth development, and tumorigenesis. Marek’s disease (MD), caused by an avian alpha-herpesvirus Marek’s disease virus (MDV), is characterized with lymphomas and immunosuppression. In this research, we hypothesize that mitochondria may play roles in response to MDV infection. To test it, mitochondrial DNA (mtDNA) abundance and gene expression in immune organs were examined in two well-defined and highly inbred lines of chickens, the MD-susceptible line 72 and the MD-resistant line 63. We found that mitochondrial DNA contents decreased significantly at the transformation phase in spleen of the MD-susceptible line 72 birds in contrast to the MD-resistant line 63. The mtDNA-genes and the nucleus-genes relevant to mtDNA maintenance and transcription, however, were significantly up-regulated. Interestingly, we found that POLG2 might play a potential role that led to the imbalance of mtDNA copy number and gene expression alteration. MDV infection induced imbalance of mitochondrial contents and gene expression, demonstrating the indispensability of mitochondria in virus-induced cell transformation and subsequent lymphoma formation, such as MD development in chicken. This is the first report on relationship between virus infection and mitochondria in chicken, which provides important insights into the understanding on pathogenesis and tumorigenesis due to viral infection.Item Allele-Specific Expression of CD4+ T Cells in Response to Marek’s Disease Virus Infection(MDPI, 2019-09-17) Bai, Hao; He, Yanghua; Ding, Yi; Carrillo, José A.; Selvaraj, Ramesh K.; Zhang, Huanmin; Chen, Jilan; Song, JiuzhouMarek’s disease (MD) is a T cell lymphoma disease induced by Marek’s disease virus (MDV), a highly oncogenic α herpesvirus primarily affecting chickens. MD is a chronic infectious disease that threatens the poultry industry. However, the mechanisms of genetic resistance for MD are complex and not completely understood. In this study, to identify high-confidence candidate genes of MD genetic resistance, high throughput sequencing (RNA-seq) was used to obtain transcriptomic data of CD4+ T cells isolated from MDV-infected and non-infected groups of two reciprocal crosses of individuals mating by two highly inbred chicken lines (63 MD-resistant and 72 MD-susceptible). After RNA-seq analysis with two biological replicates in each group, we identified 61 and 123 single nucleotide polymorphisms (SNPs) (false discovery rate (FDR) < 0.05) annotated in 39 and 132 genes in intercrosses 63 × 72 and 72 × 63, respectively, which exhibited allele-specific expression (ASE) in response to MDV infection. Similarly, we identified 62 and 79 SNPs annotated in 66 and 96 genes in infected and non-infected groups, respectively. We identified 534 and 1543 differentially expressed genes (DEGs) (FDR < 0.05) related to MDV infection in intercrosses 63 × 72 and 72 × 63, respectively. We also identified 328 and 20 DEGs in infected and non-infected groups, respectively. The qRT-PCR using seven DEGs further verified our results of RNA-seq analysis. The qRT-PCR of 11 important ASE genes was performed for gene functional validation in CD4+ T cells and tumors. Combining the analyses, six genes (MCL1, SLC43A2, PDE3B, ADAM33, BLB1, and DMB2), especially MCL1, were highlighted as the candidate genes with the potential to be involved in MDV infection. Gene-set enrichment analysis revealed that many ASE genes are linked to T cell activation, T cell receptor (TCR), B cell receptor (BCR), ERK/MAPK, and PI3K/AKT-mTOR signaling pathways, which play potentially important roles in MDV infection. Our approach underlines the importance of comprehensive functional studies for gaining valuable biological insight into the genetic factors behind MD and other complex traits, and our findings provide additional insights into the mechanisms of MD and disease resistance breeding in poultry.