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Microbial cell-free DNA (mcfDNA) sequencing is a promising tool to identify infectious pathogens when traditional methods fail to identify the causative agent. We performed a retrospective observational cohort study to evaluate clinical outcomes among pediatric and adult patients who underwent mcfDNA testing. 127 mcfDNA tests were reviewed from 112 patients. Baseline characteristics included 61 (54.5 %) adults, 52 (40.9 %) tests were from female patients, and 67 (52.8 %) tests were obtained from patients designated as immunocompromised. Of all tests obtained, 59 (46.4 %) were deemed clinically relevant. 41 (32.3 %) of tests also led to a change in antimicrobial management for the corresponding patient. No statistically significant association was ascertained between patient-specific factors and clinically relevant test results. Testing in certain clinical scenarios or high-risk settings may be useful, however further studies are needed to assess the cost-benefit of this approach.
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Ácidos Nucleicos Libres de Células , Enfermedades Transmisibles , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Femenino , Estudios Retrospectivos , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Masculino , Adulto , Niño , Persona de Mediana Edad , Adolescente , Ácidos Nucleicos Libres de Células/sangre , Preescolar , Adulto Joven , Enfermedades Transmisibles/diagnóstico , Enfermedades Transmisibles/tratamiento farmacológico , Enfermedades Transmisibles/microbiología , Anciano , Lactante , Técnicas de Diagnóstico Molecular/métodos , Anciano de 80 o más AñosRESUMEN
INTRODUCTION: Neutrophil extracellular traps (NETs) contribute to trauma-induced coagulopathy. We aimed to develop a murine multiple-injury model that induces thrombo-inflammatory response, that is, NETosis and accelerated thrombin generation. METHODS: Wild-type male mice (n = 10, aged 8-12 weeks) underwent multiple injuries (gastrocnemius crush, femur fracture, and laparotomy) and were compared with an uninjured control group (n = 10). Mice were euthanized by cardiac puncture performed 3 hours after injury. Whole blood samples were immediately processed to platelet poor plasma for thrombin generation kinetics (calibrated automated thrombogram), myeloperoxidase (MPO), and von Willebrand factor quantification. Immunohistochemistry of lung tissue was performed to assess for citrullinated histone 3 (CitH3) and MPO. A NETosis cluster was defined as 3+ neutrophils staining for CitH3 at 400× magnification (CitH3 cluster). Data were presented either as mean (SD) or median (interquartile range) with p < 0.05 significant. Sham and trauma treated animals were compared by the two-sample Wilcoxon rank-sum test. RESULTS: Animals subjected to multiple injuries had accelerated thrombin generation compared with controls with greater peak height (61.3 [41.2-73.2] vs. 28.4 [19.5-37.5] nM, p = 0.035) and shorter time to peak (3.37 [2.81-3.81] vs. 4.5 [4.08-4.75] minutes, p = 0.046). Markers of neutrophil activation were greater following multiple injuries than in controls (MPO, 961.1 [858.1-1116.8] vs. 481.3 [438.0-648.9] ng/mL; p = 0.004). NETosis, as evidenced by the aforementioned defined number of CitH3 clusters in the lung, was greater in multiple-injury animals than in controls (mean [SD], 3 [2.9] vs. 0.2 [0.7]; p = 0.009). CONCLUSION: This is the first study to demonstrate that NETosis and accelerated thrombin generation can be induced using a murine multiple-injury model, as early as 3 hours following injury.
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Traumatismo Múltiple , Trombosis , Masculino , Ratones , Animales , Tromboinflamación , Inflamación , Trombina , Neutrófilos , HistonasRESUMEN
BACKGROUND: The mammalian oviduct is a complex, fibromuscular organ known for its role in orchestrating a series of timely and dynamic changes to suitably support early embryogenesis. Climate change-induced heat stress (HS) is one of the largest single stressors compromising reproductive function in humans and farm animals via systemic changes in the redox status of the maternal environment, adversely affecting fertilization and early embryonic development. Oviductal organoids represent a unique 3-dimensional, biomimetic model to study the physiology of the oviduct and its subsequent impact on embryo development under various environmental conditions. RESULTS: Our study is the first to demonstrate an innovative approach to understanding the cascade of molecular changes sustained by bovine oviductal organoids under HS and the subsequent maternal signals harnessed within their secreted extracellular vesicles (EVs). Transcriptomic analysis of oviductal organoids exposed to HS revealed 2,570 differentially expressed genes (1,222 up- and 1,348 downregulated), while EV-coupled miRNome analysis disclosed 18 miRNAs with significant differential expression (12 up- and 6 downregulated) in EVs from thermally stressed organoids compared to EVs released from organoids cultured under thermoneutral conditions. Genes activated in oviductal organoids in response to thermal stress, include: COX1, ACTB, CST6, TPT1, and HSPB1, while miR-1246, miR-148a, miR21-5p, miR-451, and miR-92a represent the top highly abundant EV-coupled miRNAs released in response to HS. Pathway analysis of genes enriched in organoids exposed to thermal stress showed the enrichment of endocrine resistance, cellular senescence, and notch signaling pathways. Similarly, EV-coupled miRNAs released from thermally stressed organoids showed their potential regulation of genes involved in cellular senescence, p53 signaling, and TGF-beta signaling pathways. CONCLUSIONS: In conclusion, the cellular and extracellular response of bovine oviductal organoids to in vitro HS conditions reveal the prospective impact of environmental HS on the physiology of the oviduct and the probable subsequent impacts on oocyte fertilization and early embryo development. Future studies elucidating the potential impact of HS-associated EVs from oviductal organoids on oocyte fertilization and preimplantation embryo development, would justify the use of an organoid model to optimally understand the oviduct-embryo communication under suboptimal environments.
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Trompas Uterinas , MicroARNs , Humanos , Embarazo , Femenino , Animales , Bovinos , Trompas Uterinas/metabolismo , Multiómica , Estudios Prospectivos , Oviductos/metabolismo , MicroARNs/metabolismo , Organoides/metabolismo , Respuesta al Choque Térmico/genética , Mamíferos/metabolismoRESUMEN
The equine chorionic girdle is comprised of specialized invasive trophoblast cells that begin formation approximately 25 days after ovulation (day 0) and invade the endometrium to become endometrial cups. These specialized trophoblast cells transition from uninucleate to differentiated binucleate trophoblast cells that secrete the glycoprotein hormone equine chorionic gonadotropin (eCG; formerly known as pregnant mare serum gonadotropin or PMSG). This eCG has LH-like activity in the horse but variable LH- and FSH-like activity in other species and has been utilized for these properties both in vivo and in vitro. To produce eCG commercially, large volumes of whole blood must be collected from pregnant mares, which negatively impacts equine welfare due to repeated blood collections and the birth of an unwanted foal. Attempts to produce eCG in vitro using long-term culture of chorionic girdle explants have not been successful beyond 180 days, with peak eCG production at 30 days of culture. Organoids are three-dimensional cell clusters that self-organize and can remain genetically and phenotypically stable throughout long-term culture (i.e., months). Human trophoblast organoids have been reported to successfully produce human chorionic gonadotropin (hCG) and proliferate long-term (>1 year). The objective of this study was to evaluate whether organoids derived from equine chorionic girdle maintain physiological functionality. Here we show generation of chorionic girdle organoids for the first time and demonstrate in vitro production of eCG for up to 6 weeks in culture. Therefore, equine chorionic girdle organoids provide a physiologically representative 3D in vitro model for chorionic girdle development of early equine pregnancy.
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Gonadotropinas Equinas , Trofoblastos , Embarazo , Humanos , Caballos , Animales , Femenino , Gonadotropinas Equinas/farmacología , Diferenciación Celular , Gonadotropina Coriónica/farmacología , OrganoidesRESUMEN
Next-generation in vitro culture model systems are needed to study the reproductive pathologies that affect domestic animals. These 3D culture models more closely mimic normal physiological function to allow a greater understanding of reproductive pathology and to trial therapeutics without the welfare concerns and the increased time and cost associated with live animal research. Recent advances with in vitro cell culture systems utilizing human and laboratory animal tissues have been reported, but implementation of these technologies in veterinary species has been slower. Organoids are a physiologically representative 3D cell culture system that can be maintained long-term. By combining organoid culture with cryopreservation, a long-term, experimental model can be available for year-round application, thus bypassing seasonality and reproductive tract availability restrictions. Here we report the generation and cryopreservation of feline oviductal organoids for the first time. Optimal culture medium for the generation of feline oviductal organoids was established, and organoids were successfully cryopreserved using three different freezing media with organoids from each treatment demonstrating comparable viability, growth rate, and protein expression after thawing and culture. Feline oviductal organoids may facilitate an in vivo-like environment that, in conjunction with co-culture for in vitro maturation and in vitro fertilization, may positively influence in vitro gamete and embryo development, embryo quality, and pregnancy rates after embryo transfer in domestic and nondomestic felids. Furthermore, readily available cryopreserved feline oviductal organoids will facilitate this co-culture, which is of particular importance to endangered felid breeding programs where tissue and gamete samples are often opportunistically obtained with little or no notice.
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Procesamiento Proteico-Postraduccional , Proteómica , Humanos , Gatos , AnimalesRESUMEN
Introduction: Little is known regarding peripheral blood mononuclear cell telomere length (PBMC-TL) and response to traumatic injury. The objective of this study was to characterize the role of PBMC-TL in coagulation and clinical outcomes after injury. Methods: Plasma and buffy coats were prospectively collected from trauma patients and healthy volunteers. DNA was purified and PBMC-TL quantified by quantitative polymerase chain reaction. Thrombin generation kinetics were expressed as lag time (in minutes), peak height (in nanometers), time to peak (in minutes), and endogenous thrombin potential (in nM × min). Results are in median and quartiles [Q1, Q3]. P < 0.05 was considered significant (Wilcoxon rank sum testing). Results: Forty-two younger patients (21 [20, 22] years, 69% were male) and 39 older patients (62 [61, 64] years, 79% were male) were included. There was no significant difference in Clinical Frailty Scores between groups. Younger patients had longer total PBMC-TL (0.40 Mb [0.30, 0.49] vs. 0.29 Mb [0.23, 0.33], P < 0.001) and longer average PBMC-TL per chromosome (4.3 kb [3.3, 5.3] vs. 3.2 kb [2.5, 3.7], P < 0.001). When older patients were stratified by 50th percentile of PBMC-TL, there were no differences in thrombin generation; however, those with shorter telomeres were less likely to be discharged home (29% vs. 77%, P = 0.004). Older patients in the bottom quartile of PBMC-TL had shorter lag time (2.78 min [2.33, 3.00] vs. 3.33 min [3.24, 3.89], P = 0.030) and were less likely to be discharged home (22% vs. 90%, P = 0.006) than those in the top quartile of PBMC-TL. Multivariable logistic regression models revealed both increased age and shorter PBMC-TL to be independent predictors of discharge disposition other than home. Conclusion: In older trauma patients, shorter PBMC-TL is associated with accelerated initiation of thrombin generation and lower likelihood of being discharged to home.
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Leucocitos Mononucleares , Trombina , Humanos , Masculino , Anciano , Femenino , Alta del Paciente , Coagulación Sanguínea , TelómeroRESUMEN
Organoids are a type of three-dimensional (3D) cell culture that more closely mimic the in vivo environment and can be maintained in the long term. To date, oviductal organoids have only been reported in laboratory mice, women, and cattle. Equine oviductal organoids were generated and cultured for 42 days (including 3 passages and freeze-thawing at passage 1). Consistent with the reports in mouse and human oviductal organoids, the equine oviductal organoids revealed round cell clusters with a central lumen. Developing a 3D model of the mare oviduct may allow for an increased understanding of their normal physiology, including hormonal regulation. These organoids may provide an environment that mimics the in vivo equine oviduct and facilitate improved in vitro embryo production in equids.
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BACKGROUND: Despite potential benefits of medication therapy management (MTM) for complex pediatric patients, implementation of pediatric MTM services is rare. OBJECTIVES: To describe how a standardized pediatric MTM model identifies potential interventions and their impact on medication regimen complexity index (MRCI) scores in children with medical complexity (CMC) and polypharmacy. METHODS: This retrospective proof-of-concept study included pediatric patients receiving primary care in a large outpatient primary care medical home for CMC within a tertiary freestanding children's hospital from August 2020 to July 2021. Medication profiles of established patients aged 0-18 years with at least 5 active medications at the time of the index visit were assessed for medication-related concerns, potential interventions, and potential impact of proposed interventions on MRCI scores. RESULTS: Among 100 patients, an average of 3.4 ± 2.6 medication-related concerns was identified using the pediatric MTM model. Common medication-related concerns (>25% of patients) included inappropriate or unnecessary therapy, suboptimal therapy, undertreated symptom, adverse effect, clinically impactful drug-drug interaction, or duplication of therapy. A total of 97% had opportunities for 5.0 ± 2.9 potential interventions. Most common proposed interventions included drug discontinuation trial (69%), patient or caregiver education (55%), dosage form modification (51%), dose modification (49%), and frequency modification (46%). The mean baseline MRCI score was 32.6 (95% CI 29.3-35.8) among all patients. MRCI scores decreased by a mean of 4.9 (95% CI 3.8-5.9) after application of the theoretical interventions (P < 0.001). Mean potential score reduction was not significantly affected by patient age or number of complex chronic conditions. Potential impact of the proposed interventions on MRCI score was significantly greater in patients with higher baseline medication counts (P < 0.001). CONCLUSION: Most CMC would likely benefit from a pharmacist-guided pediatric MTM service. A standardized review of active medication regimens identified multiple medication-related concerns and potential interventions for nearly all patients. Proposed medication interventions would significantly reduce medication regimen complexity as measured by MRCI. Further prospective evaluation of a pharmacist-guided pediatric MTM service is warranted.
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Administración del Tratamiento Farmacológico , Polifarmacia , Adolescente , Niño , Preescolar , Enfermedad Crónica , Humanos , Lactante , Recién Nacido , Farmacéuticos , Estudios RetrospectivosRESUMEN
Culture model systems that can recapitulate the anatomy and physiology of reproductive organs, such as three-dimensional (3D) organoid culture systems, limit the cost and welfare concerns associated with a research animal colony and provide alternative approaches to study specific processes in humans and animals. These 3D models facilitate a greater understanding of the physiological role of individual cell types and their interactions than can be accomplished with traditional monolayer culture systems. Furthermore, 3D culture systems allow for the examination of specific cellular, molecular, or hormonal interactions, without confounding factors that occur with in vivo models, and provide a powerful approach to study physiological and pathological reproductive conditions. The goal of this paper is to review and compare organoid culture systems to other in vitro cell culture models, currently used to study female reproductive physiology, with an emphasis on the role of extracellular vesicle interactions. The critical role of extracellular vesicles for intercellular communication in physiological processes, including reproduction, has been well documented, and an overview of the roles of extracellular vesicles in organoid systems will be provided. Finally, we will propose future directions for understanding the role of extracellular vesicles in normal and pathological conditions of reproductive organs, utilizing 3D organoid culture systems.
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Vesículas Extracelulares , Organoides , Animales , Técnicas de Cultivo de Célula/métodos , Vesículas Extracelulares/metabolismo , Femenino , Modelos Biológicos , ReproducciónRESUMEN
Reproductive diseases in mares are a significant cause of subfertility and profound economic loss in the equine industry. Utilizing a 3D in vitro cell culture system that recapitulates the in vivo physiology will reduce time, cost, and welfare concerns associated with in vivo reproductive research in mares. If this 3D model is combined with effective cryopreservation, reproductive research on mares can occur year-round, which is not currently possible in this seasonal species. Endometrial organoids, 3D in vitro cell clusters that exhibit in vivo uterine physiology, have been established in mice, women, and mares. Here we report the first comprehensive assessment of cryopreservation of endometrial organoids in the domestic mare. Organoid growth rate was not affected by the type of freezing media. However, growth rate varied among non-cryopreserved controls, organoids cryopreserved at passage 0 (P0), and organoids cryopreserved at passage 3 (P3). Additionally, there was no difference in organoid viability among freezing media or freezing timepoint (passages). Furthermore, fresh and frozen-thawed organoids displayed positive immunohistochemical staining for ZO-1, which is a marker for intercellular tight junctions, and for periodic acid-Schiff staining as marker for organoid function through mucin production. Results demonstrate that equine endometrial organoids can be cryopreserved with 10% dimethyl sulfoxide with minimal detrimental effects while maintaining intercellular tight junctions (ZO-1) and secretory function. Availability of cryopreserved endometrial organoids may permit expanded research on uterine pathologies that negatively affect mare fertility and improve efficiency, reduce cost, and minimize animal welfare concerns associated with in vivo research in the domestic mare.
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Criopreservación , Organoides , Animales , Criopreservación/métodos , Dimetilsulfóxido/metabolismo , Dimetilsulfóxido/farmacología , Endometrio/fisiología , Femenino , Caballos , Humanos , Ratones , Organoides/metabolismo , ÚteroRESUMEN
Domestic dogs (Canis familiaris) may serve as a reservoir or a sentinel for infectious disease pathogens that can affect human and wildlife health. To understand the role of tick-borne diseases in rural and lesser developed regions, we investigated the prevalence of several tick-borne pathogens in communal dogs of Zimbabwe. Blood samples from 225 dogs in northwest Zimbabwe were assessed by serology for Ehrlichia canis, Anaplasma phagocytophilum and Borrelia burgdorferi, and 241 samples were assessed by polymerase chain reaction (PCR) for Ehrlichia. There was a high seroprevalence (73%) of E. canis-specific antibodies in domestic dogs in northwest Zimbabwe, but follow up analyses via PCR and genetic sequencing indicated only 7.5% of the canines were actively infected with the organism. Whilst indicating that an organism serologically related to E. canis is likely present in the region, this data also shows that the organism is currently present in a relative minority of the domestic dogs in the region. Its presence as evidenced by both serologic and PCR analysis is significant because of the 'one health' paradigm, where humans and wildlife may be affected by the exposure to this pathogen in domestic dogs.
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Enfermedades de los Perros/diagnóstico , Enfermedades por Picaduras de Garrapatas/veterinaria , Animales , Enfermedades de los Perros/sangre , Enfermedades de los Perros/epidemiología , Perros , Femenino , Masculino , Reacción en Cadena de la Polimerasa , Pruebas Serológicas/veterinaria , Enfermedades por Picaduras de Garrapatas/sangre , Enfermedades por Picaduras de Garrapatas/diagnóstico , Enfermedades por Picaduras de Garrapatas/epidemiología , Zimbabwe/epidemiologíaRESUMEN
The endometrium, the inner uterine lining, is composed of cell layers that come in direct contact with an embryo during early pregnancy and later with the fetal placenta. The endometrium is responsible for signals associated with normal reproductive cyclicity as well as maintenance of pregnancy. In the mare, functionally competent in vitro models of the endometrium have not been successful. Furthermore, the ability to study various reproductive processes in vitro may allow critical evaluation of signaling pathways involved in the reproductive diseases of animals that cannot be handled frequently, such as various wildlife species. Here we report the establishment of organoids, 3D structures, derived from fresh and frozen-thawed equine endometrium (Equus ferus caballus and E. f. przewalskii). Although organoids from domestic mares responded to exogenous hormonal stimuli, organoids from Przewalski's horse failed to respond to exogenous hormones. The present study represents a 'first' for any large animal model or endangered species. These physiologically functional organoids may facilitate improved understanding of normal reproductive mechanisms, uterine pathologies, and signaling mechanisms between the conceptus and endometrium and may lead to the development of novel bioassays for drug discovery.
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Endometrio/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Hormonas/farmacología , Organoides/efectos de los fármacos , Animales , Animales Salvajes , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Endometrio/metabolismo , Receptor alfa de Estrógeno/genética , Receptor alfa de Estrógeno/metabolismo , Femenino , Caballos , Organoides/metabolismo , Receptores de Progesterona/genética , Receptores de Progesterona/metabolismoRESUMEN
The inner layer of the uterus, the endometrium, is responsible and necessary for many reproductive functions. Normal reproductive cyclicity, maternal recognition of pregnancy, maternal interaction with the embryo, and interaction of the reproductive tract with pathogens are dependent on the endometrium. Although most studies have been conducted in vivo using live animals, recent advances in in vitro approaches could facilitate future research in a laboratory setting with minimal effect on animals. Many reproductive studies have been performed in vivo and in vitro in equids, but new in vitro methods to study the endometrium of mares remain unexplored. In this review, there is a description of the normal anatomy and physiology of the mare endometrium in vivo, in vitro endometrial cell culture techniques that have been previously described for the mare, and opportunities for future reproductive research using in vitro methods.
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Técnicas de Cultivo de Célula/veterinaria , Endometrio/anatomía & histología , Endometrio/fisiología , Caballos/anatomía & histología , Caballos/fisiología , Animales , Endometrio/citología , Ciclo Estral/fisiología , Femenino , EmbarazoRESUMEN
Dama gazelles (Nanger dama mhorr) are considered critically endangered by the IUCN Red List of Threatened Species since the wild population is comprised of fewer than 200 animals. Several zoos in Europe and some private ranches in the US (Texas) maintain this species in captivity as a hedge against extinction. A routine reproductive exam on an adult male revealed bilateral cysts in the testicular parenchyma. The fluid in the cysts contained copious, moderately progressive motile spermatozoa. Following examination, the gazelle was diagnosed with bilateral intratesticular spermatoceles. Electroejaculation yielded 1.5 ml of semen containing 387 × 106 total sperm with 50% motility and 45% morphologically normal sperm. The spermatoceles did not appear to cause pain or dysfunction, so no treatment was performed at this time. Since fewer than 10 animals are managed in captivity in the US, no intervention (castration/hemicastration) was considered.
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Assisted reproduction of endangered equids, such as Persian onagers (Equus hemionus onager), is vital for species conservation. Little is known about Persian onager reproductive functions, including functions of the uterine endometrium. Recently, successful cryopreservation of the domestic mare endometrium was reported, but there is no information on cryo-sensitivity or in vitro culture of endometrial tissues of any non-domestic equid. In the present study, endometrial explants from Persian onagers were cryopreserved and cultured in vitro for 5 days. There was no difference between endometrial explants when 10% and 20% dimethyl sulfoxide (DMSO) was used for cryopreservation. Cell viability and structural integrity were comparable to fresh tissue. Abundance of estrogen receptor-α (ESR1) and progesterone receptor (PGR) mRNA transcript in endometrial explants was less in most treatment groups compared to the fresh tissue control. There was variation in E-cadherin mRNA abundance in endometrial explants among treatment groups with some treatment groups having a lesser abundance compared to the control group. The abundance of Ki67 mRNA transcript of endometrial explants was not different among treatment groups compared to the control group. Results indicate that DMSO is a suitable cryoprotectant for the Persian onager endometrium, and in vitro culture in a liquid-gas interface can maintain Persian onager endometrial explants for as long as 5 days. Findings allow for a greater understanding of reproductive mechanisms in vitro for this endangered species and other domestic equids including donkeys.
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Criopreservación/veterinaria , Endometrio , Equidae/fisiología , Técnicas de Cultivo de Tejidos/veterinaria , Conservación de Tejido/veterinaria , Animales , FemeninoRESUMEN
A six-year-old multiparous Angus cow was presented for dystocia. Vaginal and rectal examinations revealed an approximately 360° counterclockwise uterine torsion. The torsion was corrected by rolling the cow counterclockwise (three episodes) with the aid of a plank coupled with manual detorsion via the vagina. The placement of obstetric chains followed by manual traction ultimately delivered a stillborn male calf with evidence of vertebral aplasia, arthrogryposis, and abdominal organ herniation. Patient history and subsequent parentage verification revealed that the calf was the result of a consanguineous (mother to son) mating. Tissue samples from the affected calf and blood samples from the dam, sire, and ten half siblings were collected for genetic testing and parentage verification. Necropsy, radiographic, and computed tomography examinations all supported a diagnosis of perosomus elumbis. Perosomus elumbis is a congenital abnormality of unknown origin(s), and this is the first report of a case associated with a consanguineous mating.
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BACKGROUND: Domestic dogs (Canis familiaris) have the potential to act as disease reservoirs for wildlife and are important sentinels for common circulating pathogens. Therefore, the infectious disease seroprevalence among domestic dogs in northern Botswana may be indicative of pathogen exposure of various wildlife species. The objective of this study was to assess the seroprevalence of Ehrlichia spp., Borrelia burgdorferi, Anaplasma spp., Dirofilaria immitis, canine adenovirus, canine parvovirus, and canine distemper virus in domestic dogs as proxies of disease prevalence in the local wildlife in the Okavango Delta region of Botswana. Statistical analysis assessed crude and factor-specific seroprevalence proportions in relation to age, sex, and geographical location as predictors of seropositivity. Logistic regression was used to identify adjusted predictors of seropositivity for each of the pathogens of interest. RESULTS: Samples from 233 dogs in a total of seven locations in Maun, Botswana, and surrounding villages were collected and serologically analyzed. No dogs were seropositive for B. burgdorferi, while low seroprevalence proportions were observed for Anaplasma spp. (2.2%) and D. immitis (0.9%). Higher seroprevalence proportions were observed for the tick-borne pathogen Ehrlichia spp. (21.0%), and 19.7% were seropositive for canine adenovirus (hepatitis). The highest seroprevalence proportions were for canine parvovirus (70.0%) and canine distemper virus (44.8%). The predictors of seropositivity revealed that adults were more likely to be seropositive for canine adenovirus, canine distemper virus, and canine parvovirus than juveniles, and location was a risk factor for canine adenovirus, canine distemper virus, canine parvovirus, and Ehrlichia spp. CONCLUSIONS: Results indicate that increasing tick control and vaccination campaigns for domestic dogs may improve the health of domestic animals, and potentially wildlife and humans in the Okavango Delta since viral and vector-borne bacterial pathogens can be transmitted between them.
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Anaplasmosis/epidemiología , Dirofilariasis/epidemiología , Moquillo/epidemiología , Enfermedades de los Perros/epidemiología , Ehrlichiosis/veterinaria , Enfermedad de Lyme/veterinaria , Infecciones por Parvoviridae/veterinaria , Anaplasma/aislamiento & purificación , Anaplasma/patogenicidad , Anaplasmosis/microbiología , Anaplasmosis/transmisión , Animales , Anticuerpos Antibacterianos/sangre , Anticuerpos Antihelmínticos/sangre , Anticuerpos Antivirales/sangre , Vectores Arácnidos/microbiología , Borrelia burgdorferi/aislamiento & purificación , Borrelia burgdorferi/patogenicidad , Botswana/epidemiología , Dirofilaria immitis/aislamiento & purificación , Dirofilaria immitis/patogenicidad , Dirofilariasis/microbiología , Dirofilariasis/transmisión , Moquillo/microbiología , Moquillo/transmisión , Virus del Moquillo Canino/aislamiento & purificación , Virus del Moquillo Canino/patogenicidad , Enfermedades de los Perros/microbiología , Enfermedades de los Perros/transmisión , Perros , Ehrlichia/aislamiento & purificación , Ehrlichia/patogenicidad , Ehrlichiosis/epidemiología , Ehrlichiosis/microbiología , Ehrlichiosis/transmisión , Femenino , Humanos , Enfermedad de Lyme/epidemiología , Enfermedad de Lyme/microbiología , Enfermedad de Lyme/transmisión , Masculino , Infecciones por Parvoviridae/epidemiología , Infecciones por Parvoviridae/microbiología , Infecciones por Parvoviridae/transmisión , Parvovirus Canino/aislamiento & purificación , Parvovirus Canino/patogenicidad , Mascotas/microbiología , Mascotas/parasitología , Mascotas/virología , Estudios Seroepidemiológicos , Garrapatas/microbiologíaRESUMEN
Availability of viable frozen-thawed endometrial tissues could facilitate detailed studies into physiologic and disease processes influencing the endometrium. This study was designed to investigate the cryosurvival of equine endometrial tissue. Previous studies in the human and horse have focused on cryopreservation of dissociated endometrial cells. To our knowledge, there are no studies on cryopreservation of endometrial explants. Our objectives were to 1) determine the influence of differing concentrations of the permeating cryoprotectant dimethyl sulfoxide (Me2SO) on viability, structural integrity, and gene expression of cryopreserved equine endometrial tissues prior to and following a 5-day explant culture in vitro and 2) examine the influence of low (1000â¯mg/L dextrose) vs high (4500â¯mg/L dextrose) glucose medium during in vitro culture. Both 10% and 20% (v/v) concentrations of Me2SO maintained viability following cryopreservation and in vitro culture. In addition, gene expression remained unaltered following cryopreservation with either 10% or 20% Me2SO. However, tissue structural integrity was slightly reduced compared to the fresh control. Furthermore, there was no difference in structural integrity, cell viability, or gene expression between low and high glucose medium during in vitro culture. Although E-cadherin and Ki67 gene expression was not different among fresh, 10% Me2SO, and 20% Me2SO treatments prior to or following tissue culture, estrogen receptor-α and progesterone receptor gene expression were reduced in all groups after explant culture. This is the first report of successful cryopreservation of equine endometrial explants.
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Criopreservación/veterinaria , Crioprotectores/farmacología , Dimetilsulfóxido/farmacología , Endometrio/fisiología , Caballos/fisiología , Animales , Supervivencia Celular , Criopreservación/métodos , Femenino , HumanosRESUMEN
We report here the first whole-genome sequences for 3 strains of Staphylococcus pseudintermedius (112N, 113N, and 114N) isolated in Africa. Samples of this opportunistic pathogen were collected from nasal swabs obtained from healthy carrier dogs in Botswana. The sequence information will facilitate spatial phylogenetic comparisons of staphylococcal species and other bacteria at the genome level.