Metabolites, Metabolic Hormones and Hematological Profiles in Mountain Goats Before the Breeding Season and During the First Trimester of Pregnancy

Objectives were to evaluate the relationships among energy-related metabolites, hormones, and hematological variables in mountain goats (Oreamnos Americans) before the breeding season and during the 1st trimester of pregnancy. Does were from herds in the Palisades (PAL) and NE Yellowstone (NEY) areas. Samples were collected from July to Aug. (before breeding season) and mid-Jan. (1st trimester of pregnancy). Sera was assayed for insulin (I), thyroxine (T4), triiodothyronine (T3), b-OH-butyrate (bOHB), blood urea nitrogen (BUN), and total protein (TP). Concentrations of TP did not differ (P > 0.05) between pregnant (P) and non-breeding season (NB) does. bOHB, I, BUN, and T3 concentrations, and the T3:T4 ratios were greater (P < 0.05) in NB does than in P does. Whereas, T4 concentrations were greater (P < 0.05) in P does than in NB does. Obviously, NB does have a different profiles of metabolites, metabolic hormones and select hematological variables compared to P does.  In conclusion, these differences may be related to P does utilizing and partitioning nutrients to support placental and fetal growth and development. These differences may also be related to the effect of season, since there were no non-pregnant does were sampled in Jan. Another factor that may be important for interpretation of these differences is location. All NB does were sampled in the PAL, while all P does were sampled in the NEY

The Intertwined Successional Development of the Lamb Gut Microbiota And Immune System (Poster)

Gastrointestinal tract (GIT) microbes play critical roles in host nutrition, health and immunological development. For adult ruminants, GIT-dwelling microbes provide ~70% of daily energy requirements. The GIT also houses 70 % of the animal’s immune system in the form of the Gut-associated Lymphatic Tissue (GALT), which houses 80% of all plasma cells and depends on microbial stimulation for maturation. Because nutrition and disease are two major factors in the economic sustainability of livestock production, our group set out to characterize the successional development of GIT microbiota and immune activity. Blood and GIT samples were collected from lambs immediately at birth through one-year of age, and from the dam’s vagina, mouth, and rectum at parturition. Blood samples were profiled for serum titers of IgM, IgA and IgG, while microbiota were profiled in GIT samples by 16S rRNA gene sequencing. Lamb GIT microbiota initially resembled the dam’s vaginal microbiota but following exposure to the dam, became rapidly more similar to the dam’s teat. GIT samples eventually formed stable climax communities similar to the dams around 180 days of age. This corresponded to the peak serum titers for each immunoglobin, which, aside from a peak in IgG at birth (likely colostral transfer), had gradually increased prior to this time. Immunoglobins peaked and then return to a sub peak level between 180 and 365 days. These results indicate dam vaginal microbiota have a short-lived impact on the neonatal microbiota, with the GIT microbiota going through a dynamic successional development to 180 d when immune function appears to peak

Using Nuclear Magnetic Resonance Spectroscopy to Develop Physiological Profiles for Bighorn Sheep (Poster)

This study employs new techniques using nuclear magnetic resonance (NMR) to assess the relative health, physiological condition, and reproductive function of wild bighorn sheep (Ovis canadensis) in Montana and Wyoming. Ongoing bighorn studies in Montana and the Greater Yellowstone Ecosystem are focused on herd attributes and the population dynamics which are affected by disease, climate, habitat and physiology. Indices of herd health and physiological status are typically obtained through expensive and time consuming lab assays and field measurements. Recently, NMR spectroscopy has been used to revolutionize the assessment of human metabolic health, and we expect that there is similar potential for studies of wildlife populations. Using NMR spectroscopy to assess metabolites associated with disease, nutrition and stress may eliminate the need for many traditional assays and techniques used today. NMR can be used to evaluate a large suite of metabolites associated with a variety of physiological functions from as little as 500 uL of serum or plasma. Blood samples from 242 sheep from 13 different herds were collected during the winters of 2013-14 and 2014-15 to develop a comprehensive metabolite panel for bighorn sheep. We have used a recently developed statistical program known as MetaboAnalyst™ to begin to analyze and evaluate differences in NMR metabolic profiles among herds and across the fall-winter season when nutritional and physiological stress is expected to be acute. We will be presenting the results of this preliminary study and discussing the potential for application in wildlife management

Nuclear Magnetic Resonance Spectroscopy Metabolic Profiles to Distinguish Geographically Isolated Populations of Mountain Goats

Basic physiological studies on mountain goats (Oreamnos americanus) are conspicuously lacking in the literature, and the physiology of this species is perhaps the least known of the high mountain ungulates. The objective of this study was to evaluate metabolic profiles of female mountain goats from five geographically distinct populations using Nuclear Magnetic Resonance (NMR) spectroscopy. Serum samples were collected from nannies located in Alaska in September (AK) from Glacier in August (GMT), from the Grand Tetons in November-December (GT), from NE Yellowstone in December (NEY) and from Absaroka in March (AB). Serum was extracted with acetone, dried and re-suspended in a standard NMR buffer. NMR spectra were analyzed with Chenomix™ software. Metabolites were identified and concentrations determined using the Chenomix™ database and the Human Metabolome Database. We identified 55 metabolites in the serum of mountain goats using this emerging technology. Of these 42 metabolites differed among the herds (P < 0.05). Of these 42 metabolites; creatinine, lactate and pyruvate distinguished (P < 0.05) each herd from another. Furthermore, using Principal Component Analyses of these metabolites allowed us to clearly differentiate metabolic profiles in carbohydrate, protein and lipid metabolism in nannies from these five populations. This study has the potential to enhance our understanding of how changes in nutrition, reproduction, susceptibility to disease, and survival rates drive population dynamics

Pregnancy Rates, Metabolites and Metabolic Hormones in Bighorn Sheep During and After the Breeding Season

Wildlife managers routinely draw blood and harvest serum when bighorn sheep (Ovis canadensis) and other ungulates are captured for management and research purposes.  Serum samples are routinely submitted to state livestock labs that perform a panel of assays to access exposure to a variety of important pathogens that cause disease, providing managers important insights.  Wildlife managers would also benefit from similar procedures that could provide assessments of reproduction, nutrition, and physiological status.  The objectives of this preliminary study were to evaluate pregnancy rates, energy-related metabolites and hormones among herds of Montana and Wyoming bighorn sheep during and after the breeding season in order to assess the general ‘health’ of herds. Metabolites and metabolic hormones are frequently used in domestic animals to evaluate nutrition, reproduction and energy balance, and potentially may provide the same insights in wildlife for managers. A total of 240 bighorn ewes were sampled from 13 herds between December 2014 and March 2015.  Samples were assayed for progesterone (P4) and pregnancy specific protein B (PSPBs) to assess reproductive cycling and pregnancy. Assays were also performed for non-esterified fatty acid, insulin, triiodothyronine and thyroxine which are metabolites and metabolic hormones that indicate nutritional and energy states of animals. We will be presenting the results of this preliminary study and discussing the relationship between pregnancy rates, energy-related metabolites and hormones and how they might be used to inform wildlife management

Developing Physiological Profiles using Nuclear Magnetic Resonance Spectroscopy to Inform Bighorn Sheep Management

This study employs new techniques using nuclear magnetic resonance (NMR) to assess the relative health, physiological condition, and reproductive function of wild bighorn sheep (Ovis canadensis)  in Montana and Wyoming. Ongoing bighorn studies in Montana and the Greater Yellowstone Ecosystem are focused on herd attributes and the population dynamics which are affected by disease, climate, habitat and physiology. Indices of herd health and physiological status are typically obtained through expensive and time consuming lab assays and field measurements. Recently, NMR spectroscopy has been used to revolutionize the assessment of human metabolic health, and we expect that there is similar potential for studies of wildlife populations. Using NMR spectroscopy to assess metabolites associated with disease, nutrition and stress may eliminate the need for many traditional assays and techniques used today. NMR can be used to evaluate a large suite of metabolites associated with a variety of physiological functions from as little as 500 ?L of serum or plasma. Blood samples from 242 sheep from 13 different herds were collected during the winters of 2013-14 and 2014-15 to develop a comprehensive metabolite panel for bighorn sheep. We have used a recently developed statistical program known as MetaboAnalyst™ to begin to analyze and evaluate differences in NMR metabolic profiles among herds and across the fall-winter season when nutritional and physiological stress is expected to be acute. We will be presenting the results of this preliminary study and discussing the potential for application in wildlife management

Sensitive and Real-Time Method for Evaluating Corneal Barrier Considering Tear Flow

We developed a new electrophysiological method mimicking tear flow to evaluate the epithelial tight junction of rabbit cornea quantitatively. We investigated the effect of tear flow on the corneal damage induced by ophthalmic preservatives using this method. An Ussing chamber system with Ag/AgCl electrodes was used in the electrophysiological experiment. The excised rabbit cornea was mounted in the Ussing chamber and the precorneal solution in the chamber was perfused with a peristaltic pump at the rate of human tear flow. Corneal transepithelial electrical resistance (TEER) was monitored as corneal barrier ability. In the electrophysiological method mimicking tear flow, we observed stable TEER, which rapidly decreased with benzalkonium chloride (BAC), an eye drop preservative. Using this system, we first found that 0.004% BAC decreased corneal TEER reversibly. A high concentration of BAC showed strong irreversible damage to the tight junction. The influence of BAC on corneal TEER was not only concentration-dependent but also tear flow rate-dependent. The electrophysiological method mimicking tear flow was useful to evaluate the corneal barrier quantitatively. Using this method, we clarified that the tear flow was important to protect the corneal damage induced by preservatives