Mariana M. Flores

A Retrospective Study of 586 Tumours in Brazilian Cattle

Records from 6,706 necropsy examinations of cattle performed over a 45-year period were surveyed and 586 cases of neoplasia were identified. The organ system most frequently affected by neoplastic disease (n=139 cases) was the alimentary tract. This finding was attributed to a high incidence of squamous cell carcinoma of the upper alimentary tract associated with the chronic ingestion of bracken fern (Pteridium aquilinum). This carcinogenic plant was also associated with a relatively high incidence (n=35 cases) of urinary bladder tumours (enzootic haematuria). Tumours of the alimentary tract were followed, in decreasing order of frequency, by tumours of the skin and subcutis (n=129 cases), haemopoietic tissue (n=101 cases), the eye and periorbital tissues (n=88 cases), the urinary system (n=44 cases), the female reproductive system (n=21 cases), the endocrine system (n=16 cases), the liver and pancreas (n=12 cases), the nervous system (n=6 cases), the respiratory system (n=6 cases) and the mammary gland (n=1 case). The primary anatomical location of 16 tumours was undetermined.

Intoxicação por alcaloides pirrolizidínicos em ruminantes e equinos no Brasil

Casos de intoxicacao por alcaloides pirrolizidinicos (APs) em ruminantes e equinos foram investigados retrospectivamente atraves do acesso aos arquivos de dois laboratorios de diagnostico veterinario no Sul e Nordeste brasileiro. Os dados obtidos foram comparados com aqueles retirados da literatura concernentes a surtos dessa toxicose no Brasil, onde ela e associada com a ingestao de plantas que contem APs dos generos Senecio, Crotalaria e Echium. Formas aguda e cronica da toxicose foram encontradas. A doenca aguda foi observada em associacao com a ingestao de Crotalaria retusa em ovinos e caprinos. C. retusa e Senecio spp. tambem foram responsaveis pela intoxicacao cronica em bovinos, equinos e ovinos. A intoxicacao por APs e uma importante causa de morte em animais pecuarios no Brasil. Essa e a principal causa de morte em bovinos na regiao Central do Rio Grande do Sul e uma das principais causas de morte em equinos na Paraiba. A epidemiologia, os sinais clinicos, a patologia e a importância da intoxicacao por APs sao descritos e discutidos.

Insights into the pathophysiology of iron metabolism in Pythium insidiosum infections

Pythium insidiosum causes life-threatening disease in mammals. Animals with pythiosis usually develop anemia, and most human patients are reported to have thalassemia and the major consequence of thalassemia, iron overload. Therefore, this study evaluated the iron metabolism in rabbits experimentally infected with P. insidiosum. Ten infected rabbits were divided into two groups: one groups received a placebo, and the other was treated with immunotherapy. Five rabbits were used as negative controls. The hematological and biochemical parameters, including the iron profile, were evaluated. Microcytic hypochromic anemia was observed in the infected animals, and this condition was more accentuated in the untreated group. The serum iron level was decreased, whereas the transferrin level was increased, resulting in low saturation. The level of stainable iron in hepatocytes was markedly decreased in the untreated group. A high correlation was observed between the total iron binding capacity and the lesion size, and this correlation likely confirms the affinity of P. insidiosum for iron. The data from this study corroborate the previous implications of iron in the pathogenesis of pythiosis in humans and animals.

Iron chelation therapy as a treatment for Pythium insidiosum in an animal model

OBJECTIVES Iron plays an important role in the pathogenesis of Pythium insidiosum. Human pythiosis frequently occurs in iron-overloaded thalassaemic patients and experimentally infected animals develop iron deficiency anaemia. Therefore, we sought to determine the in vitro and in vivo activities of the iron chelator deferasirox against P. insidiosum. METHODS In vitro, the MIC and minimum fungicidal concentration (MFC) values of deferasirox for 17 strains of P. insidiosum were determined in accordance with CLSI document M38-A2. In vivo studies were carried out in 20 inoculated rabbits divided into four groups: placebo, immunotherapy obtained from vortexed P. insidiosum cultures (14 day intervals), deferasirox (15 mg/kg/day) and a combination of immunotherapy and deferasirox. Five non-infected animals were used as controls. RESULTS The MIC and MFC values of deferasirox for P. insidiosum ranged from 12.5 to 50 mg/L and from 50 to 100 mg/L, respectively. Treatment with deferasirox alone ameliorated anaemia and normalized the serum iron levels and hepatic iron concentration in the animals. However, the mean lesion size, although decreased, did not differ significantly from that in the placebo group. The results of immunotherapy plus iron chelation therapy were worse than those of immunotherapy alone. Moreover, the disease spread to the lung tissue in 5 out of 10 deferasirox-treated animals. CONCLUSIONS Despite its limited in vitro and in vivo activity, deferasirox improved iron deficiency anaemia in P. insidiosum-infected rabbits. Further studies are needed to investigate the immunomodulatory properties observed in this study and the benefits and drawbacks of using iron-chelating drugs as an adjuvant therapy in pythiosis.

Influence of toxoplasmosis on acetylcholinesterase activity, nitric oxide levels and cellular lesion on the brain of mice.

The objective of this study was to investigate the activity of acetylcholinesterase (AChE), nitrite/nitrate (NOx) levels, as well as the biomarkers of cellular damage in the brain of mice experimentally infected with Toxoplasma gondii. Sixty mice were divided into two experiments: in experiment I the mice were infected with T. gondii/RH strain, while in experiment II they were infected with T. gondii, strains VEG and ME-49. Our evaluations were carried out on brain homogenized samples, assessing the AChE and glutathione reductase (GR) activities, and NOx, TBARS and AOPP levels in all the infected animals, compared with the control group. In both experiments, I and II, it was observed an increase in the activity of AChE and GR, as well as in the levels of NOx in the brain of infected mice with T. gondii. TBARS levels were increased in mice infected with the three different strains, RH, ME-49, and VEG. AOPP concentration was increased only in mice infected with the RH strain. Animals infected with the strains VEG and ME-49 showed histological lesions, associated with the presence of the parasite in the brain. Therefore, the infection by T. gondii is able to interfere in cholinesterase activity and NO levels, in association with oxidative stress and histological lesion.

Relationship between butyrylcholinesterase activity and liver injury in mice acute infected with Toxoplasma gondii

This study aimed to investigate the butyrylcholinesterase (BChE) activity in mice experimentally infected with Toxoplasma gondii during the acute phase. Twenty mice were divided in two groups with 10 animals each: group A was composed of uninfected mice while group B was formed by rodents infected with T. gondii. Five days after infection, blood was collected and serum separated, and fragments of liver and brain were obtained. In serum and liver homogenate was noted a significant reduction (P<0.05) in BChE activity in infected mice when compared with uninfected ones. In serum was observed an increase in the activity of alanine aminotransferase and urea, associated with reduction in alkaline phosphatase activity and in the levels of total protein and albumin. Histologically, there were foci of necrosis and parasites in the forms of tachyzoites and cysts, with bradyzoites in liver samples of infected animals. Based on these results, we conclude that toxoplasmosis reduces BChE activity in mice, and this alteration is probably related to the liver damage caused by the parasitism. Therefore, this enzymatic alteration can directly contribute to the pathogenesis of the disease.

E-NTPDase and E-ADA activities in rats experimental infected by Cryptococcus neoformans

Cryptococcus neoformans, the etiological agent of cryptococcosis, is an opportunistic fungal pathogen of immunocompromised individuals. The aim of this study was to evaluate the activities of E-NTPDase and E-ADA in rats experimentally infected by C. neoformans var. grubii. Adult rats (35) were divided in two groups: 18 for the control group (uninfected) (A), and 17 for the infected group (B). Each group was separated into three sub-groups (A1, A2, A3-B1, B2, B3), and samples were collected on 10, 20, and 30 days post-infection (PI). Leukocyte counts, IFN-γ, TNF-α, IgM, IgG levels, and E-NTPDase and E-ADA activities were analyzed. It was possible to observe that IgG and IgM seric levels of infected rats were significantly elevated (P<0.01) on days 10, 20 and 30 PI, as well as the levels of TNF-α and INF-γ when compared to uninfected rodents. Regarding E-NTPDase activity in lymphocytes, it was possible to observe that the ATP hydrolysis was significantly decreased on days 20 (P<0.01) and 30 PI (P<0.05), while ADP hydrolysis was significantly reduced only on day 20 PI (P<0.01) when compared with uninfected group. Seric E-ADA activity had a significant reduction (P<0.01) during all three evaluated periods when compared to the control group, while E-ADA activity in lymphocytes increased significantly (P<0.01) when compared to the group A on day 10 PI; however on days 20 and 30 PI, its activity was considerable reduced in lymphocytes of infected animals (P<0.01). Therefore, it is possible to conclude that the infection caused by C. neoformans in immunocompetent rats leads to changes in the purinergic signaling (NTPDase and E-ADA), concomitantly with an inflammatory response (increased levels of cytokines and immunoglobulins) associated with inflammatory infiltrates and histological lesions in the lung.

Influence of infection by Toxoplasma gondii on purine levels and E-ADA activity in the brain of mice experimentally infected mice.

The aim of this study was to assess the purine levels and E-ADA activity in the brain of mice (BALB/c) experimentally infected with Toxoplasma gondii. In experiment I (n=24) the mice were infected with RH strain of T. gondii, while in experiment II (n=36) they were infected with strain ME-49 of T. gondii. Our results showed that, for RH strain (acute phase), an increase in both periods in the levels of ATP, ADP, AMP, adenosine, hypoxanthine, xanthine (only on day 6 PI) and uric acid (only on day 6 PI). By the other hand, the RH strain led, on days 4 and 6 PI, to a reduction in the concentration of inosine. ME-49, a cystogenic strain, showed some differences in acute and chronic phase, since on day 6 PI the levels of ATP and ADP were increased, while on day 30 these same nucleotides were reduced. On day 60 PI, ME-49 induced a reduction in the levels of ATP, ADP, AMP, adenosine, inosine and xanthine, while uric acid was increased. A decrease of E-ADA activity was observed in brain on days 4 and 6 PI (RH), and 30 PI (ME-49); however on day 60 PI E-ADA activity was increased for infection by ME-49 strain. Therefore, it was possible to conclude that infection with T. gondii changes the purine levels and the activity of E-ADA in brain, which may be associated with neurological signs commonly observed in this disease.

Aspectos epidemiológicos e anatomopatológicos do hemagiossarcoma em cães: 40 casos (1965-2012)

Epidemiological and pathological aspects of hemangiosarcoma in dogs from the Central Region of the State of Rio Grande do Sul, Brazil. Out of the studied cases (n=40), aged dogs (72.2% of the cases) and dogs of German shepherd breed (20% of the cases) were clearly more frequently affected since in the total population of necropsied dogs in the same period (n=7,063) this age group and breed were comparatively less represented (respectively 14.6% e 10.1% of the cases). At necropsy (n=40) most tumors (92.5%) occurred as nodules and less frequently as masses (37.5%), affecting mainly the spleen (62.5%), lung (60%), liver (52.5%), peritoneum (42.5%), kidney (37.5%), brain (30%), pleura (25%), and heart (22.5%). Hemoperitoneum (42.5% of the cases) and resultant anemia (22.5% of the cases) were also observed. On histological examination (n=25), most hemangiosarcomas (84%), were, in general, well differentiated, of low grade (64%) and possessed a scant stroma (84%), although frequently (68%) focal areas with cells displaying some degree of atypia were seen. Necrosis, hemorrhage, and thrombosis were observed in all cases, however extramedullary hematopoiesis (28%) and benign angiomatous proliferation (12%) were less common findings. In all cases submitted to immunohistochemistry staining (n=24) the tumor cells displayed a finely granular positive staining when using anti-factor von Willebrand. Regarding anatomical classification, 55% of the hemangiosarcomas were considered as multicentric, 30% as primary tumors with one or more metastasis and 15% were solitary tumors. This paper discusses these results an suggests, based on the combination of affected organs, a scheme for setting apart primary hemangiosarcoma with metastasis from multicentric hemangiosarcoma, aiming to uniformize communication between pathologists regarding this tumor.

Changes in purine levels associated with cellular brain injury in gerbils experimentally infected with Neospora caninum.

The present study was carried out in order to assess the possible alterations in purine levels of brain, associated neuronal lesions in gerbils experimentally infected with Neospora caninum. For that, gerbils (Meriones unguiculatus) were inoculated with Nc-1 strain of N. caninum, composing two different experiments: Experiment I (EI) and experiment II (EII), where purine levels were measured along with the histopathologic study, on days 7 (EI), 15 and 30 (EII), post-infection (PI). As a result, it was possible to observe that the purine levels (ATP, ADP, AMP, adenosine, inosine and xanthine) in brain in EI are significantly reduced (p < 0.05), while in EII we faced a different pattern, since in the majority the purine levels were significantly increased (p < 0.05) on days 15 (ATP, AMP, adenosine, hypoxanthine and xanthine) and 30 PI (ATP, ADP, AMP, adenosine, and uric acid). Results of brain histopathology did not show histological lesion in animals of EI; however, in gerbils of EII it was possible to verify that the alterations (lesions) were more pronounced in gerbils evaluated on day 30 PI when compared to day 15 PI. Therefore, it was possible to conclude that the purine levels in brain were altered in both experiments, concomitant with the histopathological injuries observed in EII.