Menopause, hormone therapy and cognition: maximizing translation from preclinical research.

Menopause-associated and hormone-associated cognitive research has a rich history built from varied disciplines and species. This review discusses landmark rodent and human work addressing cognitive outcomes associated with varied experiences of menopause and hormone therapy. Critical variables in menopause and cognitive aging research are considered, including menopause etiology, background hormone milieu and parameters of exposure to estrogens and progestogens. Recent preclinical research has identified that menopause and ovarian hormone fluctuations across many neurobiological systems affect cognitive aging, mapping novel avenues for future research. Preclinical models provide insight into complex interdisciplinary relationships in a systematic and highly controlled fashion. We highlight that acknowledging the strengths and weaknesses for both preclinical and clinical research approaches is vital to accurate interpretation, optimal translation and the direction of future research. There is great value in collaboration and communication across preclinical and clinical realms, especially regarding reciprocal feedback of findings to advance preclinical models, improve experimental designs and enrich basic science translation to the clinic. In searching for biological mechanisms underlying the cognitive consequences of menopause and hormone therapies, it is noteworthy that clinical and preclinical scientists are grounded in the same fundamental goal of optimizing health outcomes for women across the lifespan.

Mitogenomic phylogeny of Callithrix with special focus on human transferred taxa.

BACKGROUND: Callithrix marmosets are a relatively young primate radiation, whose phylogeny is not yet fully resolved. These primates are naturally para- and allopatric, but three species with highly invasive potential have been introduced into the southeastern Brazilian Atlantic Forest by the pet trade. There, these species hybridize with each other and endangered, native congeners. We aimed here to reconstruct a robust Callithrix phylogeny and divergence time estimates, and identify the biogeographic origins of autochthonous and allochthonous Callithrix mitogenome lineages. We sequenced 49 mitogenomes from four species (C. aurita, C. geoffroyi, C. jacchus, C. penicillata) and anthropogenic hybrids (C. aurita x Callithrix sp., C. penicillata x C. jacchus, Callithrix sp. x Callithrix sp., C. penicillata x C. geoffroyi) via Sanger and whole genome sequencing. We combined these data with previously published Callithrix mitogenomes to analyze five Callithrix species in total. RESULTS: We report the complete sequence and organization of the C. aurita mitogenome. Phylogenetic analyses showed that C. aurita was the first to diverge within Callithrix 3.54 million years ago (Ma), while C. jacchus and C. penicillata lineages diverged most recently 0.5 Ma as sister clades. MtDNA clades of C. aurita, C. geoffroyi, and C. penicillata show intraspecific geographic structure, but C. penicillata clades appear polyphyletic. Hybrids, which were identified by phenotype, possessed mainly C. penicillata or C. jacchus mtDNA haplotypes. The biogeographic origins of mtDNA haplotypes from hybrid and allochthonous Callithrix were broadly distributed across natural Callithrix ranges. Our phylogenetic results also evidence introgression of C. jacchus mtDNA into C. aurita. CONCLUSION: Our robust Callithrix mitogenome phylogeny shows C. aurita lineages as basal and C. jacchus lineages among the most recent within Callithrix. We provide the first evidence that parental mtDNA lineages of anthropogenic hybrid and allochthonous marmosets are broadly distributed inside and outside of the Atlantic Forest. We also show evidence of cryptic hybridization between allochthonous Callithrix and autochthonous C. aurita. Our results encouragingly show that further development of genomic resources will allow to more clearly elucidate Callithrix evolutionary relationships and understand the dynamics of Callithrix anthropogenic introductions into the Brazilian Atlantic Forest.

Caracterização de biocerâmica de fosfatos de cálcio microestruturada em diferentes composições em ovinos / Bioceramic characterization of calcium phosphates microstructured in different composition in sheep

RESUMO: As biocerâmicas microporosas de fosfatos de cálcio e bifásicas de hidroxiapatita e fosfato tricálcico beta (HA/TCP-β) na forma de biomateriais granulados microporosos, são temas de pesquisas e se destacam como substitutos ósseos em aplicações biomédicas. As biocerâmicas bifásicas são biocompatíveis, bioativas, osteoindutoras e promovem a osteointegração, quando aplicados in vivo ou em meio simulado. Outro ponto diferencial dessas biocerâmicas está associado à capacidade de solubilidade que esses biomateriais apresentam quando aplicados em meio biológico, permitindo a liberação gradual de íons cálcio e fosfatos para o meio biológico, estes são absorvíeis e promovem a neoformação de um novo tecido ósseo. As biocerâmicas bifásicas de fosfatos de cálcio também se apresentam promissores em aplicações traumatológicas na reparação do tecido ósseo traumatizado e na liberação controlada de medicamentos, em tratamentos da estrutura óssea. O desempenho desses biomateriais como substitutos ósseos e na liberação controlada de medicamentos, estão associados, as suas características físicas, químicas, morfológicas e de superfície. O objetivo desse estudo foi realizar a caracterização morfológica, microestrutural dos biomateriais pela técnica de microscopia eletrônica de varredura (MEV), física com difratometria de raios X (DRX) e método de Arthur para determinação da porosidade aberta. A densidade teórica dos biomateriais bifásicos foi determinada pelo método teórico das concentrações bifásicas. Por fim, se realizou avaliação do comportamento da neoformação óssea e osteointegração dos diferentes biomateriais de fosfatos de cálcio em testes in vivo em ovinos. Os testes in vivo foram realizados em tíbias de ovinos com tempo de implantação de 03 meses. Os biomateriais implantados foram hidroxiapatita (HA), fosfato tricálcico-β (TCP-β) e composições bifásicas HA/TCP-β nas proporções: 80/20, 20/80, 70/30 e 30/70. Foram utilizadas 08 ovelhas mestiças Texel, com 12 meses de idade e peso médio de 30 kg (±5 kg), nas quais foram produzidos três defeitos ósseos em cada tíbia, sendo que quatro desses defeitos foram preenchidos por biomateriais, e dois por fragmentos ósseos (autoenxerto), grupo controle. Os animais foram eutanasiados aos 03 meses após a implantação dos biomateriais. Após a eutanásia, foram coletadas as tíbias para avaliação com o uso da técnica de microscopia eletrônica de varredura (MEV). Os resultados encontrados mostram que os biomateriais granulados microporosos são formados por uma morfologia irregular com tamanho de grânulos entre 200 μm e 500μm, outra constatação foi microestrutura microporosa interconectada dos biomateriais. O resultado obtido da porosidade aberta mostrou que os biomateriais apresentam porosidade superior a 68%. A densidade teórica se apresentou semelhante entre os biomateriais granulados de fosfatos de cálcio e sugerem boa capacidade de neoformação óssea para todos os biomateriais, sendo que o bifásico 20/80 apresentou absorção do biomaterial e neoformação óssea mais rápida quando comparada com os outros biomateriais avaliados neste estudo.

ABSTRACT: Microporous bioceramics of calcium phosphate and biphasic hydroxyapatite and tricalcium phosphate beta (HA/TCP- β) in the form of microporous granules biomaterials are research subjects and stand out as bone substitutes in biomedical application. The biphasic bioceramics are biocompatible, bioactive, osteoinductive and promote osseointegration when applied in vivo or through simulated. Another aspect of such differential solubility bioceramics is associated with the capacity that these biomaterials exhibit when applied in biological environment, enabling the gradual release of calcium and phosphate ions to the biological environment they are absorbable and promote neogenesis of new bone tissue. The biphasic bioceramics of calcium phosphate also have promising applications in traumatology in the repair of injured bone and controlled release of drugs in the bone structure treatments. The performance of these biomaterials as bone substitutes and controlled release of drugs, are associated, their physical, chemical, morphological and surface. The aim of this study was to make morphological, microstructural of biomaterials by the technique of scanning electron microscopy (SEM), physics with X-ray diffraction (XRD) and Arthur method for determination of open porosity. The theoretical density of biphasic biomaterials was determined by theoretical method of biphasic concentrations. Finally, we conducted evaluation of osteogenesis and osseointegration behavior of different biomaterials of calcium phosphates in vivo tests on sheep. In vivo tests were performed on tibias of sheep up to of 03 months. The biomaterials implanted were hydroxyapatite (HA), tricalcium phosphate-β (β-TCP) and biphasic compositions as HA/TCP-β in rates: 80/20, 20/80, 70/30 and 30/70. Eight crossbred Texel sheep, with 12 months of age and average weight of 30 kg (±5 kg) were used, in which were produced three bone defects in the tibia, four of these defects were filled with biomaterials, and two by bone fragments (autograft), as control group. The animals were euthanized at 03 months after implantation of biomaterials. After euthanasia, the tibias were collected for evaluation using the scanning electron microscopy technique (SEM). The results show that the microporous biomaterial granules are formed by an irregular morphology with grain size between 200 μm and 500μm, another finding was microporous interconnected microstructure of biomaterials. The result showed that the open porosity of the biomaterials exhibit porosity greater than 68%. The theoretical density was relatively similar between the granulates biomaterials of calcium phosphates and suggest good capacity of osteogenesis for all biomaterials, with the biphasic absorption of the biomaterial introduced 20/80 and more rapid bone formation when compared with other biomaterials evaluated in study.

Hydration study from different calcium phosphate biocements with microstructure and nanostructure.

Important features of biocements include easy molding and good wettability, hydration, and setting time during its application in biological tissue. Interest in calcium phosphate biocements is directly related to its characteristics of bioactivity, biocompatibility, and crystallographic similarity to bone apatite. This experimental study aimed to understand hydration behavior of calcium phosphate biocements with microstructure and nanostructure, with molar ratios Ca/P = 1.5; 1.6; 1.67; and 1.7 and hydration times of 5 and 30 min. The hydration tests were performed on the same solid/liquid ratio for the four Ca/P compositions. The morphology was observed via scanning electron microscopy and phases were identified with help from X-ray diffraction. The biocements showed similar effects of hydration and gelling for the periods of 5 and 30 min. The results show that these biocements can offer favorable wettability, hydration, and easy molding during the surgical procedure, which could be an innovation in implant fixation and bone tissue repair. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 820-827, 2017.

Sex-dependent programming effects of prenatal glucocorticoid treatment on the developing serotonin system and stress-related behaviors in adulthood.

Prenatal stress and overexposure to glucocorticoids (GC) during development may be associated with an increased susceptibility to a number of diseases in adulthood including neuropsychiatric disorders, such as depression and anxiety. In animal models, prenatal overexposure to GC results in hyper-responsiveness to stress in adulthood, and females appear to be more susceptible than males. Here, we tested the hypothesis that overexposure to GC during fetal development has sex-specific programming effects on the brain, resulting in altered behaviors in adulthood. We examined the effects of dexamethasone (DEX; a synthetic GC) during prenatal life on stress-related behaviors in adulthood and on the tryptophan hydroxylase-2 (TpH2) gene expression in the adult dorsal raphe nucleus (DRN). TpH2 is the rate-limiting enzyme for serotonin (5-HT) synthesis and has been implicated in the etiology of human affective disorders. Timed-pregnant rats were treated with DEX from gestational days 18-22. Male and female offspring were sacrificed on the day of birth (postnatal day 0; P0), P7, and in adulthood (P80-84) and brains were examined for changes in TpH2 mRNA expression. Adult animals were also tested for anxiety- and depressive- like behaviors. In adulthood, prenatal DEX increased anxiety- and depressive- like behaviors selectively in females, as measured by decreased time spent in the center of the open field and increased time spent immobile in the forced swim test, respectively. Prenatal DEX increased TpH2 mRNA selectively in the female caudal DRN at P7, whereas it decreased TpH2 mRNA selectively in the female caudal DRN in adulthood. In animals challenged with restraint stress in adulthood, TpH2 mRNA was significantly lower in rostral DRN of prenatal DEX-treated females compared to vehicle-treated females. These data demonstrated that prenatal overexposure to GC alters the development of TpH2 gene expression and these alterations correlated with lasting behavioral changes found in adult female offspring.

Translational cognitive endocrinology: designing rodent experiments with the goal to ultimately enhance cognitive health in women.

Understanding the cognitive impact of endogenously derived, and exogenously administered, hormone alterations is necessary for developing hormone treatments to support healthy brain function in women, especially during aging. The increasing number of studies in the burgeoning area of translational cognitive neuroendocrinology has revealed numerous factors that influence the extent and direction of female steroid effects on cognition. Here, we discuss the decision processes underlying the design of rodent hormone manipulation experiments evaluating learning and memory. It is noted that even when beginning with a clear hypothesis-driven question, there are numerous factors to consider in order to solidify a sound experimental design that will yield clean, interpretable results. Decisions and considerations include: age of animals at hormone administration and test, ovariectomy implementation, when to administer hormones relative to ovarian hormone loss, how and whether to monitor the estrous cycle if animals are ovary-intact, dose of hormone, administration route of hormone, hormone treatment confirmation protocols, handling procedures required for hormone administration and treatment confirmation, cognitive domains to be tested and which mazes should be utilized to test these cognitive domains, and control measures to be used. A balanced view of optimal design and realistic experimental practice and protocol is presented. The emerging results from translational cognitive neuroendocrinology studies have been diverse, but also enlightening and exciting as we realize the broad scope and powerful nature of ovarian hormone effects on the brain and its function. We must design, implement, and interpret hormone and cognition experiments with sensitivity to these tenets, acknowledging and respecting the breadth and depth of the impact gonadal hormones have on brain functioning and its rich plasticity. This article is part of a Special Issue entitled Hormone Therapy.

An update on the cognitive impact of clinically-used hormone therapies in the female rat: models, mazes, and mechanisms.

In women, ovarian hormone loss associated with menopause has been related to cognitive decline. Hormone therapy (HT) may ameliorate some of these changes. Understanding the cognitive impact of female steroids, including estrogens, progestogens, and androgens, is key to discovering treatments that promote brain health in women. The preclinical literature has presented elegant and methodical experiments allowing a better understanding of parameters driving the cognitive consequences of ovarian hormone loss and HT. Animal models have been a valuable tool in this regard, and will be vital to future discoveries. Here, we provide an update on the literature evaluating the impact of female steroid hormones on cognition, and the putative mechanisms mediating these effects. We focus on preclinical work that was done with an eye toward clinical realities. Parameters that govern the cognitive efficacy of HT, from what we know thus far, include but are not limited to: type, dose, duration, and route of HT, age at HT initiation, timing of HT relative to ovarian hormone loss, memory type examined, menopause history, and hormone receptor status. Researchers have identified intricate relationships between some of these factors by studying their individual effects on cognition. As of late, there is increased focus on studying interactions between these variables as well as multiple hormone types when administered concomitantly. This is key to translating preclinical data to the clinic, wherein women typically have concurrent exposure to endogenous ovarian hormones as well as exogenous combination HTs, which include both estrogens and progestins. Gains in understanding the parameters of HT effects on cognition provide exciting novel avenues that can inform clinical treatments, eventually expanding the window of opportunity to optimally enhance cognition and brain health in aging women. This article is part of a Special Issue entitled Hormone Therapy.

Recovery after chronic stress within spatial reference and working memory domains: correspondence with hippocampal morphology.

Chronic stress results in reversible spatial learning impairments in the Morris water maze that correspond with hippocampal CA3 dendritic retraction in male rats. Whether chronic stress impacts different types of memory domains, and whether these can similarly recover, is unknown. This study assessed the effects of chronic stress with and without a post-stress delay to evaluate learning and memory deficits within two memory domains, reference and working memory, in the radial arm water maze (RAWM). Three groups of 5-month-old male Sprague-Dawley rats were either not stressed [control (CON)], or restrained (6 h/day for 21 days) and then tested on the RAWM either on the next day [stress immediate (STR-IMM)] or following a 21-day delay [stress delay (STR-DEL)]. Although the groups learned the RAWM task similarly, groups differed in their 24-h retention trial assessment. Specifically, the STR-IMM group made more errors within both the spatial reference and working memory domains, and these deficits corresponded with a reduction in apical branch points and length of hippocampal CA3 dendrites. In contrast, the STR-DEL group showed significantly fewer errors in both the reference and working memory domains than the STR-IMM group. Moreover, the STR-DEL group showed better RAWM performance in the reference memory domain than did the CON group, and this corresponded with restored CA3 dendritic complexity, revealing long-term enhancing actions of chronic stress. These results indicate that chronic stress-induced spatial working and reference memory impairments, and CA3 dendritic retraction, are reversible, with chronic stress having lasting effects that can benefit spatial reference memory, but with these lasting beneficial effects being independent of CA3 dendritic complexity.

Galantamine effects on memory, spatial cue utilization, and neurotrophic factors in aged female rats.

Galantamine is an acetylcholine esterase inhibitor that has been approved for use in Alzheimer's disease. However, even though clinical studies indicate efficacy in attenuating some of the symptoms associated with the disease, there are a paucity of studies evaluating the effects of galantamine administration on cognitive performance and brain parameters in aged rats. Further, because all previous animal studies using galantamine have been performed in male rats, there is no information on how females respond to galantamine treatment. Therefore, we studied the effects of 0.3, 0.6, and 1.2 mg/kg/day galantamine in 20-month-old female rats in terms of performance on the working and reference memory water radial arm maze task. Galantamine did not influence maze performance. Furthermore, a probe trial procedure to determine extra-maze cue utilization while solving the water radial arm maze established that aged female rats utilized extramaze cues, and that they did not rely on a nonspatial chaining strategy to locate hidden platforms. Galantamine treatment had no effect on use of extramaze cues or chaining. In addition, there were no significant changes in neurotrophin levels in the frontal cortex, entorhinal cortex, hippocampus, or basal forebrain after galantamine administration. Therefore, the data reported here suggest that aged animals do utilize spatial strategies for solving a working memory task, but galantamine has no appreciable effects on this task, at least not at the doses tested.

Intraluminal irradiation for TIPS stenosis: preliminary results in a swine model.

PURPOSE: To evaluate the ability of 32phosphorus intraluminal irradiation to reduce pseudointimal hyperplasia in a transjugular intrahepatic portosystemic shunt (TIPS). MATERIALS AND METHODS: TIPS were successfully placed in 11 swine with normal portal pressures. Six animals received 15.2 Gy intraluminal irradiation to the hepatic parenchyma and venous outflow tract at the time of TIPS placement with use of a NA32P-filled balloon angioplasty catheter. Five control animals underwent TIPS and balloon angioplasty with saline. All animals were followed up for 28 days, at which time percutaneous portography was performed, the animals were killed, and the tissue around the TIPS stent was processed for histologic analysis. Maximum pseudointimal hyperplasia as a percentage of estimated TIPS diameter was calculated for each animal. RESULTS: At the time of euthanasia, all five control TIPS and all but one irradiated TIPS were occluded. Histologic analysis demonstrated considerable variability in the degree of pseudointimal hyperplasia within each TIPS and between animals. No statistically significant difference was found in the maximum pseudointimal hyperplasia, measured as a percentage of stent radius, between control (80.2%+/-17.4%) and irradiated animals (69.2%+/-25.2%). CONCLUSIONS: Irradiation of TIPS with 15.2 Gy 32P delivered at the time of TIPS placement did not significantly improve TIPS patency or reduce the degree of pseudointimal hyperplasia in swine with normal portal pressures.

Gadolinium-enhanced breathhold three-dimensional time-of-flight renal MR angiography in the evaluation of potential renal donors.

PURPOSE: To evaluate the utility of gadolinium-enhanced three-dimensional (3D) time-of-flight (TOF) magnetic resonance angiography (MRA) of the renal arteries in the evaluation of potential renal donors. MATERIALS AND METHODS: Fifty consecutive patients underwent gadolinium-enhanced 3D-TOF MRA of the renal arteries as part of their evaluation as possible renal donors. All imaging was performed on a 1.5-T system with use of a torso phased-array coil. Conventional T1-weighted axial spin-echo and T2-weighted axial fast spin-echo imaging was performed to evaluate the renal parenchyma. Coronal dynamic MRA was performed during bolus injection of 40 mL of gadolinium with use of a 3D-TOF sequence requiring a breathhold of approximately 30 seconds. Maximum-intensity-projection reconstructions were obtained of the renal arterial and venous anatomy. All studies were prospectively evaluated by a single radiologist experienced with body MRA. Intraoperative findings were used as the reference standard in 35 patients. To evaluate interobserver variability, each examination was evaluated for image quality, renal artery number, and anatomy by two radiologists experienced with MRA and blinded to the other's interpretations and surgical results. RESULTS: Ninety-eight percent of all MRAs were graded as diagnostic quality (Kappa value = 0.38; P < .05). Multiple renal arteries were identified in 29 (29%) of 100 kidneys. Four of 50 patients studied (8%) had renal parenchymal abnormalities identified with MR imaging. Sensitivity and specificity for accessory renal artery detection was 71% and 95%, respectively. Overall, accuracy for MRA in determining renal artery number was 90%. CONCLUSION: Gadolinium-enhanced breathhold 3D-TOF renal MRA is sufficient to assess the renal arteries in potential donors.

The role of percutaneous needle biopsies in the original diagnosis of lymphoma: a prospective evaluation.

PURPOSE: A prospective evaluation to determine if percutaneous needle biopsy yields enough tissue to establish the diagnosis of lymphoma and initiate therapy. PATIENTS AND METHODS: Lymphoma was diagnosed in 43 patients for the first time. Patients underwent either a core needle biopsy (n = 41) or an aspiration-type needle biopsy (n = 2) performed with imaging guidance. Immunochemical studies were performed on specimens from 39 of 43 patients (91%); flow cytometry was performed formed on specimens from 10 patients (23%). Patient progress was followed to see if biopsy results were used as a basis for treatment or if additional material was obtained with an open surgical procedure. RESULTS: On the basis of treatment decisions, needle biopsy results were sufficient for a diagnosis to be made in 36 of 43 patients (84%). In seven patients (16%), needle biopsy specimens were considered nondiagnostic, suspicious for lymphoma, or insufficient. None of the 43 patients responded to therapy in a manner that suggested the diagnosis of lymphoma to be incorrect. CONCLUSION: Image-guided needle biopsy should be the first procedure performed in the diagnosis of lymphoma, except in easily accessible superficial neck, inguinal, and axillary nodal sites.

Outbreak of vesicular dermatitis among horses at a midwestern horse show.

Dermatitis consisting of blisters on the nose and other parts of the body was reported among horses at a Midwestern horse show. Some horses also had jaundice, hematuria and anorexia. An outbreak investigation was initiated, and of 239 horses for which information could be obtained, 58 (24%) were found to have been affected. Median duration of illness was 5 days, and all horses recovered. Age, sex, water source, grain source, and stabling location were not associated with illness. The use of wood shavings bedding obtained at the show grounds was the factor most strongly associated with the development of vesicular lesions. Horses that became ill were 43 times more likely to have been bedded on wood shavings obtained from the show grounds than were horses that did not become ill. Among horses bedded on shavings from the show grounds, the risk was further increased by a factor of 5 if the shavings had been wetted. Neither organic nor heavy metal toxicants were identified in the samples of the wood shavings. However, samples did contain plant tissues originating from a tree belonging to the family Simaroubaceae, some species of which are known to cause vesicular eruptions in people.

Thyroid biopsies: fine-needle aspiration biopsy versus spring-activated core biopsy needle in 102 patients.

PURPOSE: Use of the single-action spring-activated core biopsy needle was compared with the fine needle aspiration biopsy (FNAB) technique in ultrasound (US)-guided thyroid biopsies. PATIENTS AND METHODS: Results in 102 patients who underwent sonographically guided thyroid biopsies with both fine needles and core biopsy needles were prospectively evaluated. Results from the 21-gauge FNAB (n = 102) were compared with results from 18-gauge (n = 1), 19-gauge (n = 2), 20-gauge (n = 43), or 21-gauge (n = 56) single-action spring-activated core biopsy needles. RESULTS: If the suggestive and diagnostic results were combined, the diagnostic accuracy for the core biopsy needle was 90.2% (92 of 102) and was 80.3% (82 of 102) for FNAB (P = .048). The diagnostic accuracy for the combination of both needles was 97.1% (99 of 102). The complication rate was 0.98% (n = 1). This complication was a minor soft-tissue infection successfully treated with orally administered antibiotics. CONCLUSION: In summary, sonographically guided biopsies of the thyroid performed with single-action core biopsy needles are safe and effective. The results with use of these needles are better than the results of FNAB, but the best results are obtained when both needles are used in the same patient.

Experimental equine leukoencephalomalacia, toxic hepatosis, and encephalopathy caused by corn naturally contaminated with fumonisins.

A study to evaluate the effects of dietary fumonisin B1 was conducted using 6 ponies (4 test and 2 control). A ration naturally contaminated with fumonisin B1 was fed in 3 phases: 1) 44 ppm fumonisin B1, 2) less than 1 ppm fumonisin B1, and 3) 88 ppm fumonisin B1. All ponies were monitored daily, weighed weekly, and limit fed at a rate of 0.8% body weight plus hay. Feed intake was measured daily, and a serum chemistry panel was completed once or twice weekly. Four to 7 days after initiation of the trial (Phase 1), all 4 test ponies had decreased feed consumption, and selected serum chemistry parameters were markedly elevated. On day 9, 1 pony died acutely with mild encephalopathy and hepatic necrosis. Another pony, euthanized on day 45, also had mild encephalopathy and hepatic necrosis. The remaining 2 test ponies continued the 44 ppm fumonisin B1 diet for 98 days. Phase 2 consisted of a diet with < 1 ppm fumonisin B1 for 120 days. During this phase, the serum chemistry values of the 2 ponies returned to normal. Following Phase 2, the 2 ponies were fed a diet containing 88 ppm fumonisin B1. After 75 days, 1 animal died of equine leukoencephalomalacia with mild hepatic necrosis. On day 78, the remaining pony was euthanized after showing distress; it also had leukoencephalomalacia and hepatic lesions.

Experimental reproduction of ELEM. A study to determine the minimum toxic dose in ponies.

An experiment to gain insight into the minimum toxic dose of fumonisins was conducted by feeding ponies rations with known fumonisin concentrations. Naturally contaminated corn screenings (CS) were blended with pellets, corn, and molasses to formulate individual daily diets. One group of 4 ponies was fed a ration with fumonisin B1 (FB1) varying from less than 1 ppm to 22 ppm. A second group of 5 ponies was fed a ration at varying rates containing 8 ppm FB1 for 180 days. A panel of clinical chemistry parameters was evaluated twice weekly for both groups. One pony in the first group died of equine leukoencephalomalacia (ELEM) after 225 days of which the final 55 days' diet contained 22 ppm FB1. Approximately 9 days prior to death, this animal experienced elevated liver chemistry values. All 5 ponies in the second group experienced mild, transient, clinical signs; were euthanized at 180 days; and had mild, histopathological brain lesions.

Characterization of an epizootic of pulmonary edema in swine associated with fumonisin in corn screenings.

In 1989, corn screenings were associated with acute interstitial pulmonary edema, hydrothorax, and death in swine. Attack rate was 5-50%, case fatality rate was 50-90%, and clinical course was 1-2 days. Screenings from farms with pigs affected with pulmonary edema contained 20-330 micrograms fumonisin B1 per gram. Screenings containing 92 micrograms fumonisin B1 per gram fed to weanling pigs caused pulmonary edema and death. Sterilized corn inoculated with Fusarium moniliforme and diluted 1:1 with clean corn contained fumonisin B1 (17 micrograms/g) and caused acute pulmonary edema when fed for 5 days. Survivors developed subacute hepatotoxicosis with individual hepatocellular necrosis, hepatomegalocytosis, and increased numbers of mitotic figures. Similar liver lesions occurred in pigs given fumonisin B1 intravenously at 0.8 mg/kg body weight for 14 days.

Fumonisin B1 concentrations in feeds from 45 confirmed equine leukoencephalomalacia cases.

During the fall of 1989 and winter of 1990, numerous reports of equine leukoencephalomalacia (ELEM) occurred from many regions of the United States. Typically, horses were consuming feed partially or entirely composed of corn and/or corn screenings. From October 1989 through May 1990, samples from 55 confirmed or suspected ELEM cases were received at the National Veterinary Services Laboratories, Ames, Iowa, for fumonisin B1 analysis. Samples from 9 cases in 1984-1985 were also obtained. Fumonisin B1, a mycotoxin produced by Fusarium moniliforme, causes ELEM, but little is known of naturally occurring toxic or safe levels in feeds. To determine what levels of fumonisin B1 in feeds are associated with ELEM, 45 selected cases were studied. The fumonisin B1 concentrations ranged from less than 1 ppm to 126 ppm, with the majority of the samples above 10 ppm. All types of feeds were included: corn, screenings, sweet feeds, and commercially pelleted rations. The length of exposure varied from 7 to greater than 35 days. Horse feed samples not associated with ELEM were also collected and analyzed. None of the nonproblem feed samples contained fumonisin B1 levels greater than 8 ppm.

Concentrations of fumonisin B1 in feeds associated with animal health problems.

Ninety-eight samples of feeds associated with 44 cases of equine leukoencephalomalacia (ELEM) and 83 samples of feed associated with 42 cases of a porcine pulmonary edema syndrome (PPE) were analyzed for fumonisin B1 (FB1). For comparison purposes, 51 feed samples not associated with PPE or ELEM were also analyzed. Feed associated with ELEM contained FB1 ranging from less than 1 microgram/g to 126 micrograms/g with 75% of the cases having at least 1 sample above 10 micrograms/g. Feeds associated with PPE ranged from less than 1 microgram/g to 330 micrograms/g with 71% of the cases having at least 1 sample greater than 10 micrograms/g. Quantitation was by high performance liquid chromatography (HPLC)/fluorescence using the fluorescamine derivative with confirmation by thin layer chromatography (TLC) and/or gas chromatography/mass spectroscopy (GC/MS).