Study on NGF and VEGF during the equine perinatal period

Part 1 of the study aims to: evaluate NGF and VEGF levels obtained at parturition from mare, foal and umbilical cord vein plasma, as well as in amniotic fluid; evaluate NGF and VEGF content in plasma of healthy foals during the first 72 h of life; evaluate NGF and VEGF levels at parturition in relation to selected mares’ and foals’ clinical parameters; evaluate the relationship between the two trophic factors and thyroid hormone levels in the first 72 h of life; assess mRNA expression of NGF, VEGF and BDNF and their cell surface receptors in the placenta. Part 2 aims to clinically characterize a population of foals spontaneously affected by Neonatal Encephalopathy (NE), and then to: evaluate NGF and VEGF levels in plasma samples obtained in the affected population at parturition from mare’s jugular vein, umbilical cord vein and foal’s jugular vein, as well as in amniotic fluid; evaluate NGF and VEGF content in plasma of foals affected by NE during the first 72 h of life/hospitalization; evaluate NGF and VEGF levels at birth/admission in relation to selected mares’ and foals’ clinical parameters; evaluate the relationship between the two trophic factors and thyroid hormone levels in the first 72 h of life/hospitalization; assess the mRNA expression of NGF, VEGF and BDNF, and their cell surface receptors, in the placenta of mares that delivered affected foals. The close relationship between the two trophic factors in foal plasma over time and their fine expression in placental tissues under physiological conditions appear to be key regulators of fetal development and adaptation. Their less pronounced decrease in compromised foals compared to healthy ones, their relationship with thyroid hormones over time, and the reduced expression of NGF and BDNF in placental tissues, could be key regulators in the mechanisms of NE.

Colic surgery in the equine patient: novel biomarkers, postoperative management, microbiotal influence and epidemiological analysis on the italian territory

The aim of this work was to investigate novel diagnostic and prognostic tools, postoperative treatments and epidemiologic factors impacting the outcome of surgical cases of colic. To make a more accurate diagnosis and establish a prognosis, several biomarkers have been investigated in colic patients. In this study we evaluated peritoneal PCT and blood ADMA and SDMA in SIRS positive and negative colic patients to be used as prognostic biomarkers. Our results highlighted the limits of these biomarkers in detection and the lack of specificity. In fact PCT was not detectable and even if ADMA and SDMA significantly increased in colic horses, they are not diagnostic nor prognostic markers for SIRS. Fluid therapy has been described to be crucial for the outcome of colic patients, nevertheless no guidelines have been established. Overhydration was the common practice in post surgical management. We compared cases with an extended fluid therapy protocol and cases with a restricted protocol. Results showed that survival rate and postoperative complications were similar between the groups, despite costs being significantly lower in the restricted group. The possible correlation between intestinal microbiota and colics has gained interest. In this study, cecal and colonic content from horses undergoing laparotomy were collected, and the microbiota analized. Results showed some differences in microbiota between discharged and non discharged patients, and between strangulating and non strangulating types of colic, that might suggest some influence of hind gut microbiota on the disease. A multicentric study involving three veterinary teaching hospitals on the italian territory was conducted investigating factors affecting postoperative survival and complications in colics. Results showed that the influence of age, PCV, TPP, blood lactate, reflux, type of disease, type of lesion, presence of anastomosis, duration of surgery and surgeons, were in line with literature. Amount of crystalloids used could affected the outcome.

Uso de levedura em equinos alimentados com dietas compostas de fenos de diferentes qualidades nutricionais

Com o objetivo de avaliar o efeito do uso de probiótico (Saccharomyces cerevisiae) em dietas para cavalos, utilizaram-se quatro equinos machos com média de 400 kg de peso corporal, em delineamento em quadrado latino 4 õ 4. As dietas foram compostas de concentrado e feno de capim-tifton 85 (de baixa qualidade) ou feno de alfafa (de alta qualidade), com ou sem a adição de levedura. Cada período experimental teve duração de 36 dias, com coleta total de fezes, para determinação dos coeficientes de digestibilidade aparente dos nutrientes, e coleta de fezes, para determinação do pH e da microbiologia fecal. A utilização de probiótico não alterou os coeficientes de digestibilidade aparente dos nutrientes das dietas, com exceção do extrato etéreo. A qualidade nutricional do feno influenciou os coeficientes de digestibilidade da matéria seca (MS), da proteína bruta (PB) e do extrato etéreo (EE) apenas no caso das dietas com feno de capim-tifton sem probiótico e com feno de alfafa com probiótico. A utilização de cultura de levedura aumentou a população de Lactobacillus nas fezes dos animais que receberam a dieta com feno de capim-tifton com probiótico. Não houve uso de probiótico na população de Lactobacillus nas dietas com feno de alfafa (de boa qualidade). A população de Streptococcus nas fezes aumentou com suplementação de levedura, tanto na dieta com feno de capim-tifton como naquela com feno de alfafa. A dieta com capim-tifton 85 sem probiótico promoveu redução nos valores de pH fecal 14 e 17 horas após a alimentação. Considerando ainda os valores de pH fecal nesses períodos após a alimentação, as dietas suplementadas com probiótico apresentaram valores superiores àqueles obtidos sem adição de levedura.

Desenvolvimento morfológico dos ovários em fetos eqüinos sem raça definida

O objetivo deste trabalho foi caracterizar histologicamente o desenvolvimento ovariano pré-natal em fetos eqüinos sem raça definida (SRD) nos trópicos, ressaltando surgimento e quantificação de ovogônias e folículos. Foram obtidos 131 fetos em frigorífico, os quais foram mensurados quanto ao comprimento (cm) equivalente à distância cefalococcígea (CR), seguido de corte das gônadas e fixação das amostras em Bouin por 24 horas. As idades foram estimadas por regressão. Foram preparados cortes histológicos ovarianos de 7µm de espessura, os quais foram corados com hematoxilina-eosina. O surgimento de ovogônias, folículos primordiais, em crescimento e antrais foi verificado, respectivamente, em fetos com CR entre 4,1 e 6,5cm (40 a 49 dias de gestação (DG)); 9,1 e 12,0cm (60 a 69 DG); 23,1 e 29,0cm (116 a 135 DG) e 50,1 e 65,0cm (201 a 245 DG). A quantidade máxima de ovogônias, folículos primordiais, em crescimento e antrais foi observada em fetos com CR entre, 4,1 e 6,0cm (40 a 49 DG); 16,1 e 23,0cm (90 a 115 DG); 29,1 e 36,0cm (136 a 155 DG) e 50,1 e 97,0cm (201 a 320 DG), respectivamente. Os resultados sugerem que o desenvolvimento morfológico dos ovários fetais de eqüinos SRD nos trópicos é mais precoce que o observado nos relatos de literatura referentes a estudos de eqüinos de diversas raças em regiões temperadas.

Novos registros e potencial epidemiológico de algumas espécies de mosquitos (Diptera, Culicidae), no Estado do Rio Grande do Sul

INTRODUÇÃO: A vigilância entomológica tem se mostrado uma importante estratégia de monitoramento da fauna de culicídeos com vistas a predizer o risco de exposição a espécies vetoras de patógenos. Esse trabalho apresenta uma lista de mosquitos identificados pela primeira vez no Rio Grande do Sul e discute o potencial epidemiológico de algumas espécies ocorrentes no Município de Maquiné com registros em outras regiões do Estado. MÉTODOS: Os mosquitos foram coletados com aspirador de Nasci e armadilhas CDC, entre dezembro de 2006 e dezembro de 2008, em área silvestre, rural e urbana do Município de Maquiné. RESULTADOS: Foram verificadas 55 espécies, das quais 22 são registradas pela primeira vez no estado e 10 são potencialmente vetoras do vírus Saint Louis, Oropouche, Aura, Trocara, Ilhéus, Rocio, Una, West Nile e encefalite equina do leste. CONCLUSÕES: Esses dados demonstram a importância da Vigilância Entomológica como ferramenta de informação e ação para a Vigilância em Saúde.

Morphological features and vascularization study of caprine cyclic corpus luteum

Corpus luteum is a temporary endocrine gland that regulates either the estrous cycle and pregnancy. It presents extreme dependency on the adequate blood supply. This work aims to evaluate goat corpus luteum (CL) vascular density (VD) over the estrous cycle. For that purpose, 20 females were submitted to estrus synchronization/ovulation treatment using a medroxyprogesterone intra-vaginal sponge as well as intramuscular (IM) application of cloprostenol and equine chorionic gonadotrophine (eCG). After sponge removal, estrus was identified at about 72hs. Once treatment was over, female goats were then subdivided into 4 groups (n=5 each) and slaughtered on days 2, 12, 16 and 22 after ovulation (p.o). Ovaries were collected, withdrawn and weighted. CL and ovaries had size and area recorded. Blood samples were collected and the plasma progesterone (P4) was measured through RIA commercial kits. The VD was 24.42±6.66, 36.26±5.61, 8.59±2.2 and 3.97±1.12 vessels/mm² for days 2, 12, 16 and 22 p.o, respectively. Progesterone plasma concentrations were 0.49±0.08, 2.63±0.66, 0.61±0.14 and 0.22±0.04ng/ml for days 2, 12, 16 e 22 p.o, respectively. Studied parameters were affected by the estrous cycle phase. Values greater than 12 p.o were observed. In the present work we observed that ovulation occurred predominantly in the right ovary (70% of the animals), which in turn presented bigger measures than the contra lateral one. There is a meaningful relationship between the weight and size of the ovary and these of CL (r=0.87, r=0.70, respectively, p<0.05). It is possible to conclude that morphology of goat's ovaries and plasma progesterone concentration changed according to estrous cycle stages. We propose these parameters can be used as indicators of CL functional activity.

Occurrence and molecular characterization of Cryptosporidium spp. isolated from domestic animals in a rural area surrounding Atlantic dry forest fragments in Teodoro Sampaio municipality, State of São Paulo, Brazil

The aim of this study was to assess the occurrence of Cryptosporidium in domestic animals in rural properties surrounding rain forest fragments within the municipality of Teodoro Sampaio, southeastern Brazil. Conventional sucrose flotation method followed by molecular characterization of the parasites by sequencing PCR products amplified from SSU rRNA gene were used. Stool samples were collected from domestic animals raised as pets and livestock in all rural properties surrounding three forest fragments. Samples from cattle (197), equine (63), pigs (25), sheep (11), and dogs (28) were collected from 98 rural properties. The frequency of occurrence of Cryptosporidium within each animal species was 3.0% (6/197) among cattle and 10.7% (3/28) among dogs. Cryptosporidium was not detected in stool samples from equine, sheep, and pigs. All sequences obtained from the six samples of calves showed molecular identity with Cryptosporidium andersoni while all sequences from dog samples were similar to C. canis. The frequency of occurrence of Cryptosporidium in these domestic animal species was low. The absence of C. parvum in the present study suggests that the zoonotic cycle of cryptosporidiosis may not be relevant in the region studied. The presence of Cryptosporidium species seldom described in humans may be, otherwise, important for the wild fauna as these animals are a source of infection and dissemination of this protozoan to other animal species. The impact and magnitude of infection by C. andersoni in wild ruminants and C. canis in wild canids have to be assessed in future studies to better understand the actual importance of these species in this region.

Rickettsial spotted fever in capoeirão Village, Itabira, Minas Gerais, Brazil

The present study investigated the infection by spotted fever rickettsia in an endemic area for Brazilian spotted fever (BSF; caused by Rickettsia rickettsii) in Minas Gerais State, Brazil. Human, canine and equine sera samples, and Amblyomma cajennense adult ticks collected in a rural area of Itabira City, Minas Gerais State were tested for rickettsial infection. Through Immunofluorescence Assay (IFA) we demonstrated the presence of antibodies anti-R. rickettsii in 8.2%, 81.3% and 100% of the human, canine and equine sera, respectively. None of the 356 tick specimens analyzed were positive for Rickettsia by the hemolymph test or Polymerase Chain Reaction technique (PCR) for the htrA and the gltA genes. Our serological results on horses and dogs (sentinels for BSF) appoint for the circulation of a SFG Rickettsia in the study area, however in a very low infection rate among the A. cajennense tick population.

Anticorpos anti-rickettsias do grupo da febre maculosa em equídeos e caninos no norte do Estado do Paraná, Brasil

The Brazilian Spotted Fever (BSF) is a zoonosis that can be fatal if not trteated. As there are few studies of the BSF in the Paraná State, the occurrence of BSF was serologically investigated in dogs and horses by indirect immunofluorescence assay (IFA), in the North of that State, in animals that are regarded as sentinels for BSF. A total of 241 samples of sera of equine, four of asinine, and 29 of dogs were collected in nine farms of three municipalities in the North of Paraná: Arapongas, Douradina, and Umuarama. From fifteen samples of equine serum with positive titres for RIFI, eight (53.3 %) had titre of 64 against R. rickettsii, two (13.3 %) 128 against R. rickettsii, and five (33.3 %) were reactants for R. parkeri and R. rickettsii, with titres ranging from 64 to 2048, and 128 to 1024, respectively. The results showed that domestic animals, sentinels for BSF, are under low exposition to ticks infected with spotted fever group Rickettsia, indicating low risk of human infection by these agents in the studied area.

Efeito da inclusão de diferentes fontes lipídicas e óleo mineral na dieta sobre a digestibilidade dos nutrientes e os níveis plasmáticos de gordura em equinos

Objetivou-se avaliar os efeitos da inclusão de gordura de origem animal ou vegetal e de óleo mineral sobre a aceitabilidade, a digestibilidade dos nutrientes e as concentrações plasmáticas de triglicérides e colesterol em equinos. Utilizaram-se quatro potros de 13 a 16 meses de idade recebendo dieta contendo feno de gramínea e concentrado, em delineamento quadrado latino, analisado por contrastes ortogonais. A aceitabilidade não foi influenciada pela quantidade nem pelo tipo de óleo adicionado às dietas. Os menores valores de digestibilidade de matéria seca (MS), matéria orgânica (MO) e extrato etéreo (EE) foram observados para as dietas acrescidas de óleo mineral (58,90; 60,29 e 32,02%) em comparação à dieta controle, cujos valores foram de 62,58; 64,41 e 77,72%. O coeficiente de digestibilidade do EE obtido com a dieta com óleo mineral foi menor (32,02%) que o obtido para as dietas com gordura animal (90,26%) e gordura vegetal (86,47%). A dieta com óleo mineral reduziu a concentração de HDL-C (68,75 mg/dL) em relação à dieta controle (76,00 mg/dL). A adição de fontes lipídicas e óleo mineral não influencia na aceitabilidade da dieta por equinos. O óleo vegetal não se diferencia da gordura animal quanto à digestibilidade dos nutrientes, no entanto, essas fontes de lipídios afetam a digestibilidade do extrato etéreo. A adição de óleo mineral reduz os níveis plasmáticos de HDL-C, ao passo que a adição de gordura animal e gordura vegetal não altera as concentrações plasmáticas de colesterol.

Possible Relationship Between Performance and Oxidative Stress in Endurance Horses

The purpose of this study was to evaluate oxidative stress, antioxidant biomarkers, and performance during a multiday 210-km endurance race. Nine endurance athlete horses participated in this study. Samples were always taken at the same times of day, before the beginning of the race and after every day of competition. Analytic measurements included glutathione reductase (GR) and catalase activity, thiobarbituric acid-reactive substances (TBARs), and reactive carbonylated derivatives. Competition intensity was low, with an average speed of 12.56 +/- 0.9 km/h. Four horses were unable to finish the race because of metabolic problems or fatigue. GR activity increased progressively (P < .001) throughout the competition, and TBARs showed a significant rise compared with baseline values (P < .01) but remained at the same levels throughout the 3 days of competition. Catalase and reactive carbonylated derivatives did not show any significant alterations in any time period. The best performance was obtained from horses who demonstrated higher GR capacity and/or lower TBAR concentration. In conclusion, redox. status seems to modulate horses` performance in endurance races, but further Studies are needed to better determine the adequate oxidant/antioxidant ratio to acquire optimal performance.

Pharmacokinetics of tetracycline in plasma, synovial fluid and milk using single intravenous and single intravenous regional doses in dairy cattle with papillomatous digital dermatitis

The purpose of this study was to compare the pharmacokinetics of tetracycline in plasma, synovial fluid, and milk following either a single systemic intravenous (i.v.) injection or a single i.v. regional antibiosis (IVRA) administration of tetracycline hydrochloride to dairy cattle with papillomatous digital dermatitis (PDD). To this end, plasma and synovial fluid tetracycline concentrations were compared with the minimal inhibitory concentration (MIC) values of the major bacteria, which are known to cause digital diseases and thus assess its efficacy in PDD. Residual tetracycline concentrations in milk from cows treated by both methods were also determined. Twelve Holstein cows with various stages of PDD were randomly assigned to two groups of six animals. Group 1 received a single systemic i.v. injection of 10 mg/kg of tetracycline hydrochloride. Group 2 received 1000 mg of tetracycline hydrochloride by IVRA of the affected limb. Blood, synovial fluid and milk samples were taken prior to tetracycline administration (time 0 control), and then at 22, 45 and 82 min, and 2, 3, 4, 6, 8, 12, 24, 48, 72, 96, and 120 h following drug administration. Tetracycline concentrations were determined by high-performance liquid chromatography. Mean tetracycline plasma and milk concentrations in Group 1 were higher than Group 2. The opposite was observed for synovial fluid concentrations. Group 2 synovial fluid concentrations were higher than the MIC value over 24 h for the bacteria most frequently responsible for claw disease. Compared with i.v. administration, IVRA administration of tetracycline produced very high synovial fluid and low plasma and milk concentrations.

Localisation of gelatinase activity in epidermal hoof lamellae by in situ zymography

In situ gelatin zymography is a technique, which utilises a gelatin-based emulsion overlay to detect and, more importantly, localise the gelatinase activity in underlying tissue. Gelatinase A [matrix metalloproteinase-2 (MMP-2)] and gelatinase B [matrix metalloproteinase-9 (MMP-9)] are present in equine hoof homogenates and supernatants from cultured hoof explants by SDS-PAGE gelatin zymography, and it has been assumed that the enzymes are derived solely from matrix and epithelia and not from other sources such as leucocytes. Using in situ zymography, gelatinases are shown to be localised within the equine epidermal hoof lamellae and, more specifically, are apparently produced by epidermal basal and/or parabasal cells. The pattern of expression correlates with that expected based on the progression of pathological changes observed during the onset of laminitis, thus providing further evidence that laminitis pathology probably arises as a result of inadequate local MMP regulation.

The soybean concentrated extract proliferates the vagina of adult rats

Objective: The aim of this study was to evaluate changes induced on the vagina of ovariectomized rats after treatment with soybean concentrated extract or conjugated equine estrogens and the association of both drugs. Methods: We conducted an experimental study with 50 ovariectomized rats that were randomly divided into five equal groups of 10 animals: GI received vehicle, GII received soybean concentrated extract 46 mg/kg per day, GIII received soybean concentrated extract 120 mg/kg per day, GIV received conjugated equine estrogens 50 mu g/kg per day, and GV received conjugated equine estrogens 50 mu g/kg and soybean concentrated extract 46 mg/kg per day. The substances were administered by gavage during 21 consecutive days. After that, the animals were killed under anesthesia and the vagina was removed for histological and immunohistochemical analysis. Data were initially submitted to analysis of variance. Whenever a significant difference was detected, the study was complemented with the Tukey-Kramer test for multiple comparisons. Results: GII did not show any differences on the vaginal epithelium or collagen compared with GI. GIII presented an increase in vaginal epithelium and collagen amount. GIV had the highest amount of collagen and the signals of vaginal proliferation. GV did not show any additional effect compared with GIV. Conclusions: Our data suggest that a high dose of isoflavone-rich soy extract may have positive effects on the vaginal structures of ovariectomized rats, but this action is less than that of estrogen treatment on vaginal thickness. In addition, soy extract may not block the estrogen effect on vaginal tissue.

Influence of the CYP17 polymorphism on vasomotor symptoms in postmenopausal women: a pilot study

Objective To evaluate the influence of CYP17 polymorphism on menopausal symptoms after estrogen treatment. Methods A total of 130 women were recruited, but only 100 of these were selected according to inclusion and exclusion criteria; they were treated with 0.3 mg/day conjugated equine estrogens. One year later, the study was completed by 71 women. The analysis of the Kupperman menopausal index symptoms was made with information provided by the patients on daily diary cards. Blood samples were analyzed and the women were divided into two groups based on the CYP17, 5`-untranslated region: group A (wild-type homozygote and heterozygote) and group B (mutated homozygote). Results The values for the Kupperman menopausal index were similar in both groups at baseline. The symptoms in both groups decreased after 1 year of treatment when compared to those at baseline. The improvement rate was approximately 27.09% and 32.18%, in groups A and B, respectively. The levels of estrogen after treatment were higher in both groups in comparison with the baseline values. The testosterone level rose in group B with the 1-year treatment (0.48 +/- 0.16 ng/ml), reaching a higher level than the level in group A after treatment. The sex hormone binding globulin (SHBG) level showed a significant increase after the 1-year treatment in group B, surpassing both the baseline and the after-treatment values in group A (p < 0.01). Conclusion Our data suggest that the CYP17 polymorphism did not influence the action of estrogen on menopause symptoms during the 1-year treatment. The extra production of estrogen and androgen may have been countered by the elevation of SHBG levels.