I estimated survival, cause-specific mortality, and neonatal habitat use of elk (Cervus elaphus nelsoni) calves in eastern Kentucky. I also measured habitat characteristics of elk parturition sites and annual calf production . Radio-collared females were fitted with vaginal implant transmitters and monitored for parturition behavior to locate, capture, and radio-collar calves during the springs of 2001 and 2002. Thirty-seven adult females with implant transmitters were translocated from Logan, Utah, to Addington Wildlife Management Area (WMA) in eastern Kentucky. Additional females from previous releases during 1997 and 1998 were monitored for parturition behavior. Mean calf production for all females monitored (n=77) was 66.2%. Parturition sites (n=10) were typically in closed-canopy hardwood forest within 152 m of a forest/grassland interface. Female selected sites with more boulders, andlt; 20 slope, a higher percentage of woody saplings, and thicker vegetation between 1.0 2.25 m in height compared to random sites. Twenty-seven calves were radio-collared and intensively monitored by ground and aerial telemetry. Mean annual survival was 0.766 ( 0.103). Coyote (Canis latrans) predation, meningeal worm (Parelaphostongylus tenuis), and human-caused mortality may slow population growth. Retention of implant transmitters for 40 cows ranged from 1 to 276 days (61.6 3.0). Only 2 implant transmitters worked as designed and led to calf captures. I had better success (n=25) capturing calves by monitoring parturition behavior and searching areas where pregnant cows were suspected to have given birth.

Vaginal atrophy symptoms such as dryness, irritation, and itching, are common after menopause. Vaginal estrogen therapy is the most effective treatment but not appropriate for all women. Women with estrogen-responsive breast cancer treated with aromatase inhibitor (AI) treatment, suppressing estrogen levels, often suffer from more pronounced vaginal atrophy symptoms. However, vaginal estrogen treatment is not recommended, leaving them without effective treatment options. The aim of this thesis was to study the effect of long-term anti-estrogen therapy on circulating estrogen levels and biochemical factors in vaginal mucosa in relation to morphological changes and clinical signs of vaginal atrophy. Circulating estrogen levels were analyzed by use of mass spectrometry and radioimmunoassay. Immunohistochemistry was used to study vaginal proliferation and steroid hormone receptors in vaginal mucosa. Vaginal gene expression was studied by use of microarray technology and bioinformatic tools, and validated by use of quantitative real-time PCR and immunohistochemistry. An estrogenic regulation of aquaporins and a possible role in vaginal dryness was investigated in vaginal mucosa and in Vk2E6E7 cells. Aromatase inhibitor-treated women had higher than expected estradiol and estrone levels but still significantly lower than other postmenopausal women. Aromatase was detected in vaginal tissue, the slightly stronger staining in vaginal mucosa from AI-treated women, suggest a local inhibition of vaginal aromatase in addition to the systemic suppression. Vaginal mucosa from AI-treated women had weak progesterone receptor, and strong androgen receptor staining intensity. Low estrogen levels lead to low expression of genes involved in cell adhesion, proliferation, and differentiation as well as weak aquaporin 3 protein immunostaining. The higher than expected estrogen levels in AI-treated women suggest that estrogen levels might previously have been underestimated. Systemic estrogen suppression by treatment with AIs, and possibly also by local inhibition of vaginal aromatase, results in reduced cell adhesion, proliferation, differentiation, and weak aquaporin 3 protein staining. Low proliferation and poor differentiation leads to fewer and less differentiated superficial cells affecting epithelial function and possibly also causing vaginal symptoms. Aquaporin 3 with a possible role in vaginal dryness, cell proliferation, and differentiation should be further explored for the development of non-hormonal treatment options for vaginal symptoms.

Immune activation is increasingly recognized as a critical element of HIV infection and pathogenesis, causing expansion of virus founder populations at the mucosal port of entry and eventual exhaustion of cellular immune effectors. A cohort of HIV-resistant (HIV-R) commercial sex workers (CSW) in Nairobi, Kenya, have increased levels of anti- inflammatory factors in vaginal secretions and reduced peripheral immune activation ("immune quiescence"). The mucosal immune micro-environment underlying HIV susceptibility is well-known to be influenced by concurrent sexually transmitted infections, however the role of commensal microbiota is poorly characterized. Bacterial vaginosis (BV), characterized by a shift from Lactobacillus to Gardnerella and Prevotella as dominant members of vaginal microbiota, is a risk factor for HIV acquisition in studies worldwide. However, the etiology and ecological dynamics of BV remain enigmatic, and the mechanisms by which BV increases HIV susceptibility are not fully defined. Protective functional characteristics of Lactobacillus microbiota, including acid and hydrogen peroxide (H2O2) production, may reinforce physicochemical defences of vaginal mucus, stimulate innate epithelial defences and/or modulate activation status of HIV target cells. Therefore, the goal of this study was to determine if reduced BV and increased Lactobacillus colonization are the basis for resistance to HIV in this cohort. Vaginal specimens from a group of 242 CSW were examined, including microscopic diagnosis of BV, culture-based functional analyses and phylogenetic profiling by ultra-deep sequencing. HIV-R individuals were just as likely to have BV compared to other HIV- negative (HIV-N) individuals, and no more likely to be colonized with acid- or H2O2- ii producing bacteria, however two BV-related phylotypes identified by deep sequencing were significantly more likely to be observed in HIV-N individuals (p=0.0002 and p=0.006). HIV+ individuals were significantly more likely than HIV– individuals to have E. coli detected by deep sequencing (p<0.0001) and less likely to have Lactobacillus crispatus (p=0.0006). A coherent set of differences in culture-based and culture- independent characteristics were observed in individuals with BV diagnoses compared to BV– individuals. This study has generated an unprecedented amount of information regarding the composition, structure and function of the vaginal microbiota in African CSW, fundamentally defining many aspects of BV microbiology. Elucidation of the relationship between complex microbial communities and protective mucosal responses against HIV infection should be a priority for future research.

Immune activation is increasingly recognized as a critical element of HIV infection and pathogenesis, causing expansion of virus founder populations at the mucosal port of entry and eventual exhaustion of cellular immune effectors. A cohort of HIV-resistant (HIV-R) commercial sex workers (CSW) in Nairobi, Kenya, have increased levels of anti- inflammatory factors in vaginal secretions and reduced peripheral immune activation ("immune quiescence"). The mucosal immune micro-environment underlying HIV susceptibility is well-known to be influenced by concurrent sexually transmitted infections, however the role of commensal microbiota is poorly characterized. Bacterial vaginosis (BV), characterized by a shift from Lactobacillus to Gardnerella and Prevotella as dominant members of vaginal microbiota, is a risk factor for HIV acquisition in studies worldwide. However, the etiology and ecological dynamics of BV remain enigmatic, and the mechanisms by which BV increases HIV susceptibility are not fully defined. Protective functional characteristics of Lactobacillus microbiota, including acid and hydrogen peroxide (H2O2) production, may reinforce physicochemical defences of vaginal mucus, stimulate innate epithelial defences and/or modulate activation status of HIV target cells. Therefore, the goal of this study was to determine if reduced BV and increased Lactobacillus colonization are the basis for resistance to HIV in this cohort. Vaginal specimens from a group of 242 CSW were examined, including microscopic diagnosis of BV, culture-based functional analyses and phylogenetic profiling by ultra-deep sequencing. HIV-R individuals were just as likely to have BV compared to other HIV- negative (HIV-N) individuals, and no more likely to be colonized with acid- or H2O2- ii producing bacteria, however two BV-related phylotypes identified by deep sequencing were significantly more likely to be observed in HIV-N individuals (p=0.0002 and p=0.006). HIV+ individuals were significantly more likely than HIV– individuals to have E. coli detected by deep sequencing (p<0.0001) and less likely to have Lactobacillus crispatus (p=0.0006). A coherent set of differences in culture-based and culture- independent characteristics were observed in individuals with BV diagnoses compared to BV– individuals. This study has generated an unprecedented amount of information regarding the composition, structure and function of the vaginal microbiota in African CSW, fundamentally defining many aspects of BV microbiology. Elucidation of the relationship between complex microbial communities and protective mucosal responses against HIV infection should be a priority for future research.

The vaginal microbial community is critical to a woman’s health and the health of her family. Bacterial vaginosis (BV) is a polymicrobial syndrome characterized by a shift of the vaginal microbiota from a Lactobacillus dominated community to a dense biofilm containing a complex mixture of organisms. Although BV is an important risk factor for poor reproductive health outcomes, the etiology of BV is poorly understood. Gardnerella vaginalis is a hallmark species of BV. Phylogenetic analysis of cpn60 universal target sequences from metagenomic studies of the vaginal microbiome and from G. vaginalis isolates resolved four subgroups within the species. This subdivision, supported by whole genome similarity comparisons, demonstrated that these subgroups might represent different species. Among a group of African women, only G. vaginalis subgroup B was significantly more abundant in women with BV relative to women with Nugent scores not consistent with BV. To characterize the subgroups further, several phenotypic and molecular factors of G. vaginalis subgroups were assessed. Proteomic profiles of isolates within each subgroup formed unambiguous clusters. Sialidase gene sequences were detected in all subgroups, however enzymatic activity was detected only in subgroup B. Two isolates of subgroup B isolates (N153 and N101) were incapable of growth in 7% CO2. Given the well-known relationship between an anaerobic microbiota and BV, anaerobic isolates of G. vaginalis are potentially important players in the vaginal microbial community. To determine genome content differences that could account for the phenotypic difference, whole genome sequences of four G. vaginalis subgroup B isolates representing facultative and anaerobic phenotypes were determined. Comparison of genomes led to the identification of genes predicted to encode proteins involved in cell wall biogenesis and protection from oxidative damage that might account for the observed phenotypes. The cpn60 universal target based methodology that improved resolution of the vaginal microbiota including G. vaginalis was applied in a prospective study of the vaginal microbiome of women with preterm premature rupture of membranes (PPROM). The objectives were to characterize the vaginal microbiota of women following PPROM, and to determine if microbiome composition at the time of rupture predicts latency duration and perinatal outcomes. Only 13/70 samples collected from 36 women were dominated by Lactobacillus spp., the expected profile for healthy women, while Megasphaera type 1 and Prevotella spp. were detected in all samples. Microbiome profiles at the time of membrane rupture did not cluster by gestational age at PPROM, or latency duration. Microbial profiles were unstable over the latency period, with dramatic shifts in composition between weekly samples, and an overall decrease in Lactobacillus abundance. Mollicutes were detected by PCR in 81% (29/36) of women, and these women had significantly lower gestational age at delivery and correspondingly lower birth weight infants than Mollicutes negative women. Taken together, the results presented in this thesis demonstrate the value of high resolution profiling of the vaginal microbiome using cpn60 UT sequences. The resolution of subgroups within G. vaginalis has potentially significant implications for women's health diagnostics, requiring a shift away from considering G. vaginalis as a single entity. The PPROM study provides foundational information that may lead to the identification of informative sequence patterns, providing clinicians with better tools for expectant management following PPROM.

Šio darbo uždaviniai įvertinti kalių, sergančių piometra, makšties citologinius ir kraujo parametrų pakitimus. Įvertinti amžiaus įtaką piometros pasireiškimui. Nustatyti laiką po rujos iki piometros klinikinių požymių atsiradimo. Nustatyti mirties atvejus dėl patologijos. Buvo tiriamos trys grupės kalių. I grupė – 16 piometra sergančių patelių. II grupė – 4 kalės, porujo periode, kliniškai sveikos. III grupė 4 – patelės, anestrus periode, kurioms kraujyje nustatyta leukocitozė, bet dėl kitų priežasčių nei piometra ar vaginitas. II ir III grupės – kontrolinės grupės, kurių rezultatai lyginami su I grupės. Tepinėlis iš makšties imamas Transwab (Jungtinė Karalystė) terpės pagalba. Gauta klinikinė medžiaga pernešama ant objektyvinio stiklelio citologijai ir skirtingų mitybinių terpių mikrobiologijai. Objektyvinis stiklelis dažomas May Grün – Giemza dažais. Makšties tepinėlis vertinamas mikroskopo pagalba. Atliekami uždegiminių ir epitelinių ląstelių santykio skaičiavimai. Kraujas imamas iš v. cefalica. Buvo pastebėti makšties tepinėlio citologiniai skirtumai tarp trijų tiriamųjų grupių. Uždegiminių su epitelinėmis ląstelėmis santykio vidurkis I grupei – 9,54 ± 3,62, II grupei – 0,09 ± 0,018, III grupei uždegiminių ląstelių nebuvo aptikta, todėl santykis ligus 0. Visose grupėse dominavo bazalinės ląstelės. 75 % kalių, sergančių piometra, konstatuota leukocitozė, 81,5 % granuliocitozė. 12,25 % buvo nustatyta mažakraujystė. 37,50 % gimdos uždegimu sergančioms kalėms išskirta E... [toliau žr. visą tekstą] / The aim of this study is to evaluate cytological changes in vaginal smears of bitches with pyometra, to record changes in total blood count and blood biochemistry, to identify microorganisms isolated from vagina. Other aims are to asses the average age of bitches with the diagnosed pathology; to count days that passed after estrus period and the first clinical signs of pyometra were noticed; to evaluate the rate of mortality. Three groups of animals were evaluated. The 1st one 16 bitches, diagnosed with pyometra. 2nd group 4 bitches, in diestrus period, clinically healthy. 3rd group 4 bitches diagnosed with blood leukocytosis but for other reasons than pyometra or vaginitis. 2nd and 3rd groups served as control groups. Vaginal smears and microbiological samples were taken from vagina with Transwab (UK), the obtained material was smeared on objective glass and coloured with May Grün – Giemsa for microscopical evaluation. Ratio of WBC and epithelial cells was counted. For microbiology sample was smeared on different agars for isolation of various microorganisms. Blood was drawn from v. cefalica. We observed clear cytological diferences between healthy animals and ill with pyometra. Average ratio WBC to epithelial cells for group I 9,54 ± 3,62, for group II 0,09 ± 0,018, for group III no WBC were observed so the ratio is equal to 0. In all groups basal epithelial cells were the dominant ones. 75 % bitches with pyometra had leukocytosis, 81,25 % had granuliocytosis. Anemia was... [to full text]

Diss. (sammanfattning) Linköping : Linköpings universitet, 2008. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 4 uppsatser.