Experimental oral administration of pollen beetle (Astylus atromaculatus) to cattle results in an acute lethal gastrointestinal disease.

In the summer of 2023, ingestion of Astylus atromaculatus (pollen beetle) was linked to spontaneous fatal disease in grazing cattle and sheep in Argentina and Uruguay. While the disease was experimentally reproduced in sheep and guinea pigs in the 1970's, no experimental reproductions have been attempted in cattle, and controversy exists as to whether this insect is indeed noxious to cattle and at which dose. Here, we demonstrate that A. atromaculatus causes acute fatal disease in Hereford calves at single oral dosages of 2.5, 4.5, 10.0, and 15.0 g of insect/kg body weight. Death or severe disease necessitating euthanasia occurred at 38 to 48 hours postinoculation regardless of the dose, suggesting that the single fatal dosage is likely <2.5 g/kg body weight (this dose representing approximately 850 mL of intact beetles in a 100 kg calf). Clinically, the disease was characterized by acute anorexia, prolonged recumbency, reluctance to move, listlessness/apathy, depression, ruminal hypomotility and tympany, hypothermia, bruxism with frothing at the mouth, and mucoid diarrhea progressing to death. Hematologic and biochemical alterations included hemoconcentration, stress/acute inflammatory leukogram, negative energy balance, and ketosis. The pathological hallmark of this experimental disease is acute necrotizing omaso-reticulo-rumenitis, fibrinohemorrhagic enteritis, and exfoliative colitis with intralesional chitinous insect fragments. While A. atromaculatus might contain a gastrointestinal toxin or pathogen, extensive toxicological testing failed to identify a causative toxin. Other pathomechanisms such as direct physical damage caused by insect fragments on the alimentary tract seem plausible, although further studies are needed to elucidate the pathogenesis of A. atromaculatus-associated disease.

Solar-induced dorsal skin necrosis in sheep.

BACKGROUND: Solar-induced thermal burns of dark skin over the dorsum have been reported in dogs, sheep and a pig. OBJECTIVES: This report describes an outbreak of solar-induced thermal burns over the dorsal skin of criollo and Texel sheep in Uruguay. ANIMALS AND METHODS: Cross-bred criollo and pure-bred Texel adult ewes from a flock of 80 animals presented with severe skin lesions. Eight animals were evaluated clinically and skin biopsy specimens were collected from three ewes for histopathology. Epidemiological data were collected. RESULTS: Black/brown criollo sheep presented with extensive, wide, linear dorsal skin necrosis extending from the interscapular to the lumbosacral area. Necrotic skin was firm, dry and largely detached from the underlying subcutis. Nonpigmented Texel sheep showed milder lesions with rapid re-epithelialization and healing. Histological features were consistent with third-degree burns, characterized by full-thickness coagulative necrosis of epidermis and dermis, including blood vessels and adnexa. The cumulative incidence of the disease was 21% during the 30 days post-shearing, affecting 50% of criollo and 7.4% of Texel sheep (P < 0.001). CONCLUSIONS: The findings are consistent with post-shearing, sun-induced thermal burns leading to dorsal skin necrosis. Risk factors include sunlight exposure during hot months after shearing, dark skin and obesity.

Pollen beetle (Astylus atromaculatus)-associated gastroenteric disease in cattle: report of 6 natural outbreaks.

Astylus atromaculatus is a pollen beetle native to South America, commonly found in crop flowers. Experimental intoxication of sheep and guinea pigs by this beetle resulting in fibrinonecrotizing enteritis has been reported. We describe here 6 natural outbreaks of intoxication in cattle associated with consumption of alfalfa (5 of 6) and mixed native (1 of 6) pastures heavily contaminated with A. atromaculatus. The outbreaks occurred during the summer (January-February) of 2023 in Argentina (n = 4) and Uruguay (n = 2), in beef cattle under extensive or semi-extensive rearing systems, with overall cumulative incidence and mortality of 22.3% and 17.8%, respectively. The main clinical signs included acute onset of anorexia, lethargy, hyperthermia, hindlimb weakness, reluctance to move, and diarrhea, for up to 15 d. In 2 outbreaks, sudden death was observed. Eight Hereford, Angus, and/or crossbreed heifers, cows, steers, and/or calves were autopsied. Gross and microscopic findings included multifocal necrosis with fibrinous pseudomembranes in the forestomachs and/or small and large intestines. Fragments or whole specimens of A. atromaculatus were identified in the ruminal content of all animals. Testing for multiple gastroenteric pathogens was negative as was testing of A. atromaculatus for cantharidin and batrachotoxin. GC-MS and LC-MS/MS performed on the beetles did not identify any known toxic compounds. Based on the exposure to A. atromaculatus-contaminated pasture, gross and microscopic lesions, and negative results of all testing for multiple gastroenteric pathogens, a diagnosis of intoxication by A. atromaculatus is proposed. Disease caused by A. atromaculatus consumption has not been reported previously in cattle, to our knowledge.

Spontaneous intoxication of sheep by the pollen beetle Astylus atromaculatus: 4 outbreaks in Uruguay and Argentina.

Astylus atromaculatus Blanchard is a native beetle of South America that feeds on pollen. During the summer of 2022-2023 in Argentina and Uruguay, an explosive infestation of these insects occurred in pastures in which ruminants were grazing. This was believed to be associated with a severe drought, which had significantly reduced the flowering of crops. Three farms in Uruguay and one in Argentina were visited to examine the flocks and perform autopsies. Affected sheep had watery diarrhea, anorexia, depression, and ruminal atony. The average morbidity, mortality, and case fatality rates were 7.5%, 4.3%, and 68%, respectively. The main gross findings in all animals were in the jejunum; the serosa had multifocal hemorrhages, and the mucosa was necrotic and covered by a pseudomembrane. Microscopically, the mucosa had partial-to-complete necrosis of the lamina propria, as well as loss of villus and crypt epithelium with neutrophilic infiltration. Overlying the necrotic mucosa was a pseudomembrane of fibrin, cell debris, desquamated epithelial cells, degenerate neutrophils, and bacteria. Many specimens of A. atromaculatus were in all paddocks in which sheep grazed, as well as in the ruminal content of the autopsied animals.

Aerobic co-oxidation of hemoglobin and aminoacetone, a putative source of methylglyoxal.

Aminoacetone (1-aminopropan-2-one), a putative minor biological source of methylglyoxal, reacts like other α-aminoketones such as 6-aminolevulinic acid (first heme precursor) and 1,4-diaminobutanone (a microbicide) yielding electrophilic α-oxoaldehydes, ammonium ion and reactive oxygen species by metal- and hemeprotein-catalyzed aerobic oxidation. A plethora of recent reports implicates triose phosphate-generated methylglyoxal in protein crosslinking and DNA addition, leading to age-related disorders, including diabetes. Importantly, methylglyoxal-treated hemoglobin adds four water-exposed arginine residues, which may compromise its physiological role and potentially serve as biomarkers for diabetes. This paper reports on the co-oxidation of aminoacetone and oxyhemoglobin in normally aerated phosphate buffer, leading to structural changes in hemoglobin, which can be attributed to the addition of aminoacetone-produced methylglyoxal to the protein. Hydroxyl radical-promoted chemical damage to hemoglobin may also occur in parallel, which is suggested by EPR-spin trapping studies with 5,5-dimethyl-1-pyrroline-N-oxide and ethanol. Concomitantly, oxyhemoglobin is oxidized to methemoglobin, as indicated by characteristic CD spectral changes in the Soret and visible regions. Overall, these findings may contribute to elucidate the molecular mechanisms underlying human diseases associated with hemoglobin dysfunctions and with aminoacetone in metabolic alterations related to excess glycine and threonine.

Senecio spp. transboundary introduction and expansion affecting cattle in Uruguay: Clinico-pathological, epidemiological and genetic survey, and experimental intoxication with Senecio oxyphyllus.

The genus Senecio is distributed worldwide, being responsible of poisoning in livestock and humans. Many species of Senecio have high invasion and expansion capacity, highly competitive with agricultural and native plant species, causing ecological damage. Particularly in Uruguay, poisoning by Senecio have grown exponentially to reach epidemic proportions. Herein we describe Seneciosis as a re-emerging and expanding epidemic disease affecting cattle, by describing clinico-pathological, epidemiological and genetic variation of species involved, as well as an experimental intoxication with Senecio oxyphyllus. For this, a study was carried out on 28 cattle farms in Eastern Uruguay, with history of seneciosis from 2010 to 2016. Plants of fifty populations of Senecio were sampled, in 2015 and 2016, for identification, analysis of alkaloids and study of genetic variation. In turn, post-mortem examination was performed in cattle of natural and an experimental case to confirm the intoxication, showing microscopic characteristic lesions (hepatomegalocytosis, diffuse fibrosis and ductal reaction). Four species of Senecio were identified: S. oxyphyllus, S. madagascariensis, S. selloi and S. brasiliensis. In the genetic study, 489 molecular markers of amplified sequence-related polymorphisms (SRAP), associated with species and pasture, were used for genetic variation analysis. There was no statistically significant association between genetic variation determined by molecular markers and population (specimens of same species collected from the same farm), botanically determined species, or geographical origin. The increase of seneciosis in cattle in the last years, the presence of species not identified to the moment with implication in the poisoning outbreaks and expansion of these plants shows that the disease is in an epidemic growing active stage. In turn, the experimental poisoning with S. oxyphyllus confirms its chronic hepatotoxic effect, being an emergent species for the region, of high distribution and toxic risk. This latter turned out the main Senecio species involved. This case of expansion of harmful plant for animal production and desirable plant species, can be useful as a model of ecopathological characterization, which is likely to occur with other toxic plants in different geographical ranges globally.

Lysophosphatidylcholine triggers cell differentiation in the protozoan parasite Herpetomonas samuelpessoai through the CK2 pathway.

BACKGROUND: Protozoa are distantly related to vertebrates but present some features of higher eukaryotes, making them good model systems for studying the evolution of basic processes such as the cell cycle. Herpetomonas samuelpessoai is a trypanosomatid parasite isolated from the hemipteran insect Zelus leucogrammus. Lysophosphatidylcholine (LPC) is implicated in the transmission and establishment of Chagas disease, whose etiological agent is Trypanosoma cruzi. LPC is synthesized by T. cruzi and its vectors, the hemipteran Rhodnius prolixus and Triatoma infestans. Platelet-activating factor (PAF), a phospholipid with potent and diverse physiological and pathophysiological actions, is a powerful inducer of cell differentiation in Herpetomonas muscarum muscarum and T. cruzi. The enzyme phospholipase A2 (PLA2) catalyzes the hydrolysis of the 2-ester bond of 3-sn-phosphoglyceride, transforming phosphatidylcholine (PC) into LPC. METHODS: In this study, we evaluated cellular differentiation, PLA2 activity and protein kinase CK2 activity of H. samuelpessoai in the absence and in the presence of LPC and PAF. RESULTS: We demonstrate that both PC and LPC promoted a twofold increase in the cellular differentiation of H. samuelpessoai, through CK2, with a concomitant inhibition of its cell growth. Intrinsic PLA2 most likely directs this process by converting PC into LPC. CONCLUSIONS: Our results suggest that the actions of LPC on H. samuelpessoai occur upon binding to a putative PAF receptor and that the protein kinase CK2 plays a major role in this process. Cartoon depicting a model for the synthesis and functions of LPC in Herpetomonas samuelpessoai, based upon our results regarding the role of LPC on the cell biology of Trypanosoma cruzi [28-32]. N nucleus, k kinetoplast, PC phosphatidylcholine, LPC lysophosphatidylcholine, PLA2 phospholipase A2, PAFR putative PAF receptor in trypanosomatids [65], CK2 protein kinase CK2 [16].

Aminoacetone, a putative endogenous source of methylglyoxal, causes oxidative stress and death to insulin-producing RINm5f cells.

Aminoacetone (AA), triose phosphates, and acetone are putative endogenous sources of potentially cytotoxic and genotoxic methylglyoxal (MG), which has been reported to be augmented in the plasma of diabetic patients. In these patients, accumulation of MG derived from aminoacetone, a threonine and glycine catabolite, is inferred from the observed concomitant endothelial overexpression of circulating semicarbazide-sensitive amine oxidases. These copper-dependent enzymes catalyze the oxidation of primary amines, such as AA and methylamine, by molecular oxygen, to the corresponding aldehydes, NH4(+) ion and H2O2. We recently reported that AA aerobic oxidation to MG also takes place immediately upon addition of catalytic amounts of copper and iron ions. Taking into account that (i) MG and H2O2 are reportedly cytotoxic to insulin-producing cell lineages such as RINm5f and that (ii) the metal-catalyzed oxidation of AA is propagated by O2(*-) radical anion, we decided to investigate the possible pro-oxidant action of AA on these cells taken here as a reliable model system for pancreatic beta-cells. Indeed, we show that AA (0.10-5.0 mM) administration to RINm5f cultures induces cell death. Ferrous (50-300 microM) and Fe(3+) ion (100 microM) addition to the cell cultures had no effect, whereas Cu(2+) (5.0-100 microM) significantly increased cell death. Supplementation of the AA- and Cu(2+)-containing culture medium with antioxidants, such as catalase (5.0 microM), superoxide dismutase (SOD, 50 U/mL), and N-acetylcysteine (NAC, 5.0 mM) led to partial protection. mRNA expression of MnSOD, CuZnSOD, glutathione peroxidase, and glutathione reductase, but not of catalase, is higher in cells treated with AA (0.50-1.0 mM) plus Cu(2+) ions (10-50 microM) relative to control cultures. This may imply higher activity of antioxidant enzymes in RINm5f AA-treated cells. In addition, we have found that AA (0.50-1.0 mM) plus Cu(2+) (100 microM) (i) increase RINm5f cytosolic calcium; (ii) promote DNA fragmentation; and (iii) increase the pro-apoptotic (Bax)/antiapoptotic (Bcl-2) ratio at the level of mRNA expression. In conclusion, although both normal and pathological concentrations of AA are probably much lower than those used here, it is tempting to propose that excess AA in diabetic patients may drive oxidative damage and eventually the death of pancreatic beta-cells.

Isolation of pathogenic Leptospira strains from naturally infected cattle in Uruguay reveals high serovar diversity, and uncovers a relevant risk for human leptospirosis.

Leptospirosis is a neglected zoonosis with worldwide distribution. The causative agents are spirochete bacteria of the Leptospira genus, displaying huge diversity of serovars, the identity of which is critical for effective diagnosis and vaccination purposes. Among many other mammalian species, Leptospira infects cattle, eliciting acute signs in calves, and chronic disease in adult animals often leading to abortions. In South America, and including in Uruguay, beef and dairy export are leading sources of national income. Despite the importance of bovine health, food safety, and bovine-related dissemination of leptospirosis to humans, extremely limited information is available as to the identity of Leptospira species and serovars infecting cattle in Uruguay and the South American subcontinent. Here we report a multicentric 3-year study resulting in the isolation and detailed characterization of 40 strains of Leptospira spp. obtained from infected cattle. Combined serologic and molecular typing identified these isolates as L. interrogans serogroup Pomona serovar Kennewicki (20 strains), L. interrogans serogroup Canicola serovar Canicola (1 strain), L. borgpetersenii serogroup Sejroe serovar Hardjo (10 strains) and L. noguchii (9 strains). The latter showed remarkable phenotypic and genetic variability, belonging to 6 distinct serogroups, including 3 that did not react with a large panel of reference serogrouping antisera. Approximately 20% of cattle sampled in the field were found to be shedding pathogenic Leptospira in their urine, uncovering a threat for public health that is being largely neglected. The two L. interrogans serovars that we isolated from cattle displayed identical genetic signatures to those of human isolates that had previously been obtained from leptospirosis patients. This report of local Leptospira strains shall improve diagnostic tools and the understanding of leptospirosis epidemiology in South America. These strains could also be used as new components within bacterin vaccines to protect against the pathogenic Leptospira strains that are actually circulating, a direct measure to reduce the risk of human leptospirosis.

Oxidative stress is enhanced by hypothermia imposed on cerulein-induced pancreatitis in rats.

BACKGROUND: Hypothermia is a frequent event in severe acute pancreatitis (AP) and its real effects on the normal pancreas have not been well demonstrated. Moreover, neither have its effects on the outcome of acute pancreatitis been fully investigated. One hypothesis is that oxidative stress may be implicated in lesions caused or treated by hypothermia. AIM OF THE STUDY: To investigate the effect of hypothermia in cerulein-induced acute pancreatitis (CIAP) in rats and the role played by oxidative stress in this process. METHODS: Male Wistar rats were divided into hypothermic and normothermic groups. Hypothermia was induced with a cold mattress and rectal temperature was kept at 30 masculineC for one hour. Acute pancreatitis was induced with 2 doses of cerulein (20 ìg/kg) administered at a one-hour interval. Serum amylase, pancreas vascular permeability by Evan's blue method, pancreas wet-to-dry weight ratio and histopathology were analyzed in each group. RESULTS: When compared with normothermic rats, hypothermic animals, with cerulein-induced acute pancreatitis, showed higher levels of pancreatic vascular permeability (p < 0.05), pancreas wet-to-dry weight ratio (p = 0.03), and histologically verified edema (p < 0.05), but similar serum amylase levels. The hypothermic group showed a higher oxidized-reduced glutathione ratio than the normothermic group. CONCLUSION: Moderate hypothermia produced a greater inflammatory response in established acute pancreatitis induced by cerulein in rats. Moreover, this study suggests that oxidative stress may be one of the mechanisms responsible for the worse outcome in hypothermic rats with cerulein-induced acute pancreatitis.

Tannic acid inhibits in vitro iron-dependent free radical formation.

The antioxidant activity of tannic acid (TA), a plant polyphenol claimed to possess antimutagenic and anticarcinogenic activities, was studied by monitoring (i) 2-deoxyribose degradation (a technique for OH detection), (ii) ascorbate oxidation, (iii) ascorbate radical formation (determined by EPR analysis) and (iv) oxygen uptake induced by the system, which comprised Fe(III) complexes (EDTA, nitrilotriacetic acid (NTA) or citrate as co-chelators), ascorbate and oxygen. TA removes Fe(III) from the co-chelators (in the case of EDTA, this removal is slower than with NTA or citrate), forming an iron-TA complex less capable of oxidizing ascorbate into ascorbate radical or mediating 2-deoxyribose degradation. The effectiveness of TA against 2-deoxyribose degradation, ascorbate oxidation and ascorbate radical formation was substantially higher in the presence of iron-NTA (or iron-citrate) than with iron-EDTA, which is consistent with the known formation constants of the iron complexes with the co-chelators. Oxygen uptake and 2-deoxyribose degradation induced by Fe(II) autoxidation were also inhibited by TA. These results indicate that TA inhibits OH formation induced by Fe(III)/ascorbate/O(2) mainly by arresting Fe(III)-induced ascorbate oxidation and Fe(II) autoxidation (which generates Fe(II) and H(2)O(2), respectively), thus limiting the production of Fenton reagents and OH formation. We also hypothesize that the Fe(II) complex with TA exhibits an OH trapping activity, which explains the effect of TA on the Fenton reaction.

Poisoning of sheep by Vernonia plantaginoides (Less.) Hieron in Uruguay.

Vernonia plantaginoides (Less.) Hieron, previously known as Vernonia squarrosa, is a rhizomatous subshrub with purple flowers that is prevalent in the natural grassland of Uruguay, Argentina, and southern Brazil. We report an outbreak of V. plantaginoides (yuyo moro) intoxication in sheep in Treinta y Tres Department, northeastern Uruguay. A total of 54 of 463 (12%) recently weaned lambs died 2-7 days after entering a natural pasture that had been invaded by sprouting V. plantaginoides The first cases were found dead. Affected lambs showed marked jaundice, edema of the face, ears, and eyelids, and severe photodermatitis. At the autopsies of 3 lambs, the carcass was yellow, the liver was enlarged with a marked acinar pattern ("nutmeg liver"), and hemorrhages were observed on serous membranes. Microscopic lesions were characterized by diffuse periacinar hepatocellular necrosis and cholemic nephrosis. Three female lambs were experimentally dosed with the aerial parts of V. plantaginoides collected immediately after the outbreak. The lamb that was dosed once with 40 g/kg body weight died after 36 h with severe hepatic necrosis. The lamb dosed with 20 g/kg daily for 4 days showed clinical signs and microscopic lesions in the liver with multiple apoptotic hepatocytes in the periacinar zone. The third lamb, dosed with 30, 17, and 15 g/kg daily over 3 days, respectively, showed transient clinical signs and a rise in liver enzymes, but recovered, and no lesions were found postmortem. These results demonstrate that V. plantaginoides was responsible for severe field outbreaks of poisoning in sheep in Uruguay.

Effects of linalool and eugenol on the survival of Leishmania (L.) infantum chagasi within macrophages.

The most commonly used drugs against visceral leishmaniasis are based on pentavalent antimonial compounds, which have played a fundamental role in therapy for over 70 years. However, the treatment is painful and has severe toxic side effects that can be fatal. Antimonial resistance is spreading and reaching alarming proportions. Linalool and eugenol have been shown to kill Leishmania (L.) amazonensis and Trypanosoma cruzi at low doses. In the present study, we demonstrate the effects of linalool and eugenol, components of essential oils, on Leishmania (L.) infantum chagasi, one of the causative agents of visceral leishmaniasis. We compared the effects of those compounds to the effects of glucantime, a positive control. In L. infantum chagasi killing assays, the LD50 for eugenol was 220µg/ml, and that for linalool was 550µg/ml. L. infantum chagasi was added to cultures of peritoneal mouse macrophages for four hours prior to drug treatment. Eugenol and linalool significantly decreased the number of parasites within the macrophages. Eugenol and linalool enhanced the activities of the L. infantum chagasi protein kinases PKA and PKC. Linalool also decreased L. infantum chagasi oxygen consumption. In conclusion, both linalool and eugenol promoted a decrease in the proliferation and viability of L. infantum chagasi. These effects were more pronounced during the interaction between the parasites and peritoneal mouse macrophages.

Molecular detection of Rangelia vitalii in domestic dogs from Uruguay.

The piroplasm Rangelia vitalii is the etiological agent of canine rangeliosis, a severe disease affecting domestic dogs in South America. Two domestic dogs from two different Departments (Salto and Treinta y Tres) of Uruguay presented with clinical signs such as apathy, anorexia, pale mucous membranes, jaundice, and hemorrhagic manifestations, suggestive of a canine vector-borne disease. Molecular analysis, based on PCR and DNA sequencing of portions of the 18S rRNA gene, revealed that both dogs were infected by R. vitalii. Two consensus sequences, one from Salto and one from Treinta y Tres, differed from each other by only 1 nucleotide (99.8% similarity) and were 99.8-100% identical to corresponding sequences of R. vitalii from Brazil and Argentina available in GenBank. Through phylogenetic analysis inferred by the 18S rRNA gene, the two Uruguayan sequences of R. vitalii were aligned with the corresponding sequences from 7 other R. vitalii sequences available in GenBank (5 from Brazil and, 2 from Argentina) under high bootstrap support. The two dogs of the present study were negative for Ehrlichia canis according to the E. canis-specific real-time PCR assay. Our findings not only confirm the occurrence of R. vitalii in Uruguay but also provide the southernmost record of this re-emerging agent. The only previous report of R. vitalii in Uruguay dated from 1976, a period when molecular analyses were not available. We provide the first molecular detection of R. vitalii in Uruguay. Currently, canine rangeliosis is confirmed to occur in Brazil, Argentina, and Uruguay.

Public humanization policies: integrative literature review.

The study aimed to investigate the scientific literature on Public Humanization Policies, available in online periodicals, from 2009 to 2012, in the health field. This is an integrative literature review conducted in the Virtual Health Library databases: Latin-America and Caribbean Health Sciences (Lilacs) and the Scientific Electronic Library Online (SciELO) and Portal Capes. Data were collected in July 2013. To this end, the following Health Sciences Descriptors (DeCS) were used: "Humanization of Care," "Public Policies," "National Humanization Policy". The sample consisted of 27 articles about the investigated theme. From the publications selected for the research, three categories emerged according to their respective approaches: National Human-ization Policy: history and processes involved in its implementation; National Humanization Policy: health professionals contribution; Humanization and in the care process. The study showed that the National Humanization Policy is an important benchmark in the development of health practices. For this reason, there is a pressing multiplication of related reflections on ways to promote human-ization in health services.

Aminoacetone induces loss of ferritin ferroxidase and iron uptake activities.

Aminoacetone (AA) is a threonine and glycine metabolite overproduced and recently implicated as a contributing source of methylglyoxal (MG) in conditions of ketosis. Oxidation of AA to MG, NH4+, and H2O2 has been reported to be catalyzed by a copper-dependent semicarbazide sensitive amine oxidase (SSAO) as well as by copper- and iron ion-catalyzed reactions with oxygen. We previously demonstrated that AA-generated O2*-. and enoyl radical (AA*) induce dose-dependent Fe(II) release from horse spleen ferritin (HoSF); no reaction occurs under nitrogen. In the present study we further explored the mechanism of iron release and the effect of AA on the ferritin apoprotein. Iron chelators such as EDTA, ATP and citrate, and phosphate accelerated AA-promoted iron release from HoSF, which was faster in horse spleen isoferritins containing larger amounts of phosphate in the core. Incubation of apoferritin with AA (2.5-50 mM, after 6 h) changes the apoprotein electrophoretic behavior, suggesting a structural modification of the apoprotein by AA-generated ROS. Superoxide dismutase (SOD) was able to partially protect apoferritin from structural modification whereas catalase, ethanol, and mannitol were ineffective in protection. Incubation of apoferritin with AA (1-10 mM) produced a dose-dependent decrease in tryptophan fluorescence (13-30%, after 5 h), and a partial depletion of protein thiols (29% after 24 h). The AA promoted damage to apoferritin produced a 40% decrease in apoprotein ferroxidase activity and an 80% decrease in its iron uptake ability. The current findings of changes in ferritin and apoferritin may contribute to intracellular iron-induced oxidative stress during AA formation in ketosis and diabetes mellitus.

Equine pythiosis in the eastern wetlands of Uruguay / Pitiose equina em áreas úmidas do leste do Uruguai

Equine pythiosis is an ulcerative and granulomatous disease of the skin, caused by the oomycete Pythium insidiosum (Pythiaceae). The objective of this study was to describe seven cases of equine pythiosis that occurred from 2012 to 2017 in the eastern region of Uruguay. Six of the seven cases occurred in the eastern wetland ecosystems of the Merin basin, and the remaining case occurred in the wetland fluvial plains of the Tacuarembó River. Lesions consisted of a large, rapidly growing ulcerated tumor with abundant granulation tissue, serosanguineous secretion, and fistulous tracts containing large concretions or kunkers. The animals presented intense pruritus, claudication and loss of body condition, with death or euthanasia in extremis in six cases. The main histological lesions consisted of an eosinophilic and pyogranulomatous inflammatory process, with numerous foci of eosinophilic necrosis (kunkers), collagenolysis, and a Splendore-Hoeppli reaction. In all cases, silver coloration (Grocott) showed intralesional hyphae compatible with P. insidiosum, which was confirmed by immunohistochemistry in three cases. A horse in the terminal phase of the disease was treated with triamcinolone acetonide (50mg IM every 15 days), and fully recovered after 1 year. It is concluded that equine pythiosis is prevalent in the wetland ecosystems of eastern Uruguay and that treatment with triamcinolone is auspicious.(AU)

Pitiose é uma doença granulomatosa e ulcerativa da pele dos equinos causada pelo oomyceto Pythium insidiosum (Pythiaceae). O objetivo deste trabalho foi descrever sete casos de pitiose equina que ocorreram de 2012 a 2017 na região leste do Uruguai. Seis dos sete casos ocorreram no ecossistema de áreas pantanosas da bacia da bacia da Lagoa Mirim Merin e o restante nas planícies fluviais pantanosas do rio Tacuarembó. As lesões se caracterizaram por tumores ulcerados de crescimento rápido com abundante tecido de granulação, secreção serossanguinolenta e presença de tratos fistulosos contendo material coraloide ou kunkers. Os equinos apresentavam prurido intenso, claudicação e perda da condição corporal e seis morreram ou foram eutanasiados in extremis. As principais lesões histológicas consistiam de um processo inflamatório piogranulomatoso com numerosos focos de necrose eosinofílicos (kunkers), colagenólise e reação de Splendori-Hoepli. Em todos os casos a impregnação pela prata (Grocott) revelou a presença de hifas intralesionais compatíveis com P. insidiosum, o que foi confirmado pela imuno-histoquímica em três casos. Um equino em fase terminal da doença foi tratado com triamcinolona acetonida (50mg, IM, a cada 15 dias), recuperando-se completamente após um ano. Conclui-se que a pitiose é uma enfermidade presente em áreas úmidas na região leste do Uruguai e o tratamento com triamcinolona pode ser uma alternativa promissora.(AU)

Occurrence of squamous cell carcinoma in Milchschaf sheep in Uruguay / Ocorrência do carcinoma de células escamosas em ovinos Milchschaf em Uruguai

ABSTRACT: In this study, the epidemiological and pathological features of an outbreak of squamous cell carcinoma (SCC) in adult female sheep and a young ram, Friesian Milchschaf breed or crossbreed is described. Seven adult females belonging to an original flock nucleus of 20 animals were affected, as well as a young ram belonging to the same nucleus was affected in other establishment. Multiple verrucous neoplastic masses of several months of evolution were evident in the head region, with local invasion and regional lymph node metastases. Histological studies revealed a cancerous stage of invasion of the dermis and neoplastic proliferation characteristic of invasive SCC, and a pre-cancerous stage with solar elastosis and chronic solar keratosis lesions induced by sunlight. Decrease in latitude, increased level of solar radiation with long exposure to sunlight, and the phenotypic characteristics of the breed are the main factors responsible for the high prevalence of SCC, showing the susceptibility of the Friesian Milchschaf breed and crossbreed in temperate zones as Uruguay.

RESUMO: Neste estudo descrevem-se as características epidemiológicas e patológicas de um surto de carcinoma de células escamosas (SCC) em ovelhas adultas e ram jovem, raça Friesian Milchschaf ou cruza. Foram afetadas sete fêmeas adultas pertencentes a um núcleo de rebanho original de 20 animais, bem como um ram jovem pertencente ao mesmo núcleo em que foi afetado em outro estabelecimento. Múltiplas neoplásicas verrucosas de vários meses de evolução foram evidentes na região principal, com invasão local e metástases de linfonodos regionais. Estudos histológicos revelaram um estágio cancerígeno de invasão da derme e proliferação neoplásica característica do SCC invasivo e um estágio pré-cancerígeno com elastose solar e lesões de ceratose solar crônica induzidas pela luz solar. A diminuição da latitude, o aumento do nível de radiação solar com longa exposição à luz solar e as características fenotípicas da raça são os principais fatores responsáveis pela alta prevalência de SCC, mostrando a susceptibilidade da raça Friesian Milchschaf e cruzada em zonas temperadas como o Uruguai.

A survey of Senecio spp. affecting livestock in Uruguay and their associated pyrrolizidine alkaloid content / Senecio spp. Afetando a pecuária no Uruguai e sua associação com alcaloides pirrolizidínicos

ABSTRACT: In Eastern Uruguay there has been a significant increase of seneciosis in grazing livestock with most affected localities related to counties neighboring the Brazilian border. A survey in 28 farms associated with poisoning outbreaks in grazing cattle in Eastern Uruguay was carried out. Fifty populations of Senecio plants were collected for alkaloid analysis and species identification. Four species were identified: S. oxyphyllus DC, S. madagascariensis Poir, S. brasiliensis (Spreng.) Less., and S. selloi DC. Alkaloids were identified by a combination of GC-MS and HPLC-MS analysis and included: retrorsine in S. oxyphyllus; retrorsine, usaramine, and senecivernine/senecionine in S. selloi; retrorsine, senecivernine/senecionine, integerrimine, and usaramine in S. madagascariensis; and integerrimine, retrorsine and senecionine in S. brasiliensis. Total mean alkaloid concentration was reported to be highest in S. brasiliensis (17.6mg/g) followed by S. oxyphyllus (6.2mg/g), S. selloi (1.8mg/g) and S. madagascariensis (0.6mg/g). Alkaloid concentrations were also reported to be higher in 2015 vs. 2016 probably due to a common environmental factor. The species S. oxyphyllus and S. madagascariensis were not previously recognized as toxic plants in Eastern Uruguay. Particularly, S. oxyphyllus was present in 82% of the farms surveyed and occurred in high density with relative high concentrations of pyrrolizidine alkaloids suggesting S. oxyphyllus may be the main species involved in the reported outbreaks of seneciosis.

RESUMO: No leste do Uruguai houve um aumento significativo da seneciosis no gado de pastagem com a maioria das localidades afetadas em municípios vizinhos à fronteira brasileira. Aplicou-se um questionário em 28 fazendas associadas a surtos de intoxicação em bovinos de pastagem na fronteira leste do Uruguai. Coletaram-se cinquenta populações de Senecio para análise de alcalóides e identificação de espécies. Identificaram-se quatro espécies: S. oxyphyllus DC, S. madagascariensis Poir, S. brasiliensis (Spreng.) Less. e S. selloi DC. Identificaram-se os seguintes alcalóides mediante análise combinado de GC-MS e HPLC-MS: retrorsina em S. oxyphyllus; retrorsina, usaramina e senecivernina/senecionina em S. selloi; retrorsina, senecivernina/senecionina, integerimina e usaramina em S. madagascariensis; e integerrimina, retrorsina e senecionina em S. brasiliensis. A concentração média total de alcalóides foi mais elevada em S. brasiliensis (17,6mg/g) seguido de S. oxyphyllus (6,2mg/g), S. selloi (1,8mg/g) e S. madagascariensis (0,6mg/g). As concentrações de alcaloides foram maiores em 2015 em vista à 2016, provavelmente, devido a um fator ambiental comum. As espécies S. oxyphyllus e S. madagascariensis não foram previamente reconhecidas como plantas tóxicas no leste do Uruguai. Particularmente, S. oxyphyllus esteve presente em 82% das fazendas pesquisadas e ocorreu em alta densidade com concentrações relativamente elevadas de alcalóides de pirrolizidina sugerindo que S. oxyphyllus pode ser a principal espécie envolvida nos surtos relatados de seneciosis.