<Emphasis Type="Italic">Lupinus albus</Emphasis> Conglutin Gamma Modifies the Gene Expressions of Enzymes Involved in Glucose Hepatic Production <Emphasis Type="Italic">In Vivo</Emphasis>

Lupinus albus seeds contain conglutin gamma (Cγ) protein, which exerts a hypoglycemic effect and positively modifies proteins involved in glucose homeostasis. Cγ could potentially be used to manage patients with impaired glucose metabolism, but there remains a need to evaluate its effects on hepatic glucose production. The present study aimed to analyze G6pc, Fbp1, and Pck1 gene expressions in two experimental animal models of impaired glucose metabolism. We also evaluated hepatic and renal tissue integrity following Cγ treatment. To generate an insulin resistance model, male Wistar rats were provided 30% sucrose solution ad libitum for 20 weeks. To generate a type 2 diabetes model (STZ), five-day-old rats were intraperitoneally injected with streptozotocin (150 mg/kg). Each animal model was randomized into three subgroups that received the following oral treatments daily for one week: 0.9% w/v NaCl (vehicle; IR-Ctrl and STZ-Ctrl); metformin 300 mg/kg (IR-Met and STZ-Met); and Cγ 150 mg/kg (IR-Cγ and STZ-Cγ). Biochemical parameters were assessed pre- and post-treatment using colorimetric or enzymatic methods. We also performed histological analysis of hepatic and renal tissue. G6pc, Fbp1, and Pck1 gene expressions were quantified using real-time PCR. No histological changes were observed in any group. Post-treatment G6pc gene expression was decreased in the IR-Cγ and STZ-Cγ groups. Post-treatment Fbp1 and Pck1 gene expressions were reduced in the IR-Cγ group but increased in STZ-Cγ animals. Overall, these findings suggest that Cγ is involved in reducing hepatic glucose production, mainly through G6pc inhibition in impaired glucose metabolism disorders.     

Mycoplasma hyopneumoniae colonization of pigs sired by different boars

Differences in Mycoplasma hyopneumoniae colonization were evaluated in experimentally inoculated pigs sired by 3 different boars of the same genetic line. Forty-six pigs were used, including a treatment group and positive and negative control groups. The pigs were intratracheally inoculated with an M. hyopneumoniae suspension or with Friis media as a placebo. To evaluate differences in the magnitude of colonization during a 35-day period, nasal and tracheal swabs were collected weekly and tested by nested polymerase chain reaction (N-PCR). Temperature, weight and circulating antibodies were measured for 35 days. At 11 and 35 d postinoculation the pigs were necropsied and macroscopic and microscopic lesions were determined. A section of bronchus was tested by the indirect immunofluorescence test (IFAT), scanning electron microscopy (SEM) and N-PCR. The N-PCR results from the nasal and tracheal swabs showed that the pigs sired by one boar (B3) had a distinctive colonization pattern, different from that of the pigs from the other 2 boars and from the positive controls. SEM studies demonstrated that at 35 d postinoculation a higher proportion of B3 pigs had lower numbers of mycoplasmas attached to the cilia compared with B1 and B2 offspring. No significant differences were observed in temperature and weight gain among groups by ANOVA; however, with use of a 2 × 2 table, temperature differences were observed between pigs sired by boars B1 and B2 at 4 d postinoculation. No pigs seroconverted, showed gross or microscopic lesions, or had positive IFAT results. These results provide evidence of differences in patterns of colonization between pigs sired by different boars, suggesting a possible genetic effect.     

Determination of the protease activity in a Cuban strain of Babesia bovis]

The attenuation of a Babesia bovis strain depends on its protease content. The present work evaluates this parameter on a virulent strain, before and after attenuation by quick passages on splenectomized calves. The protease activity at different pH values was determined in protein fractions from the blood of calves. The enzymatic test showed marked differences between the protease content of both substrains.     

Foot-and-mouth disease virus typing by complement fixation and enzyme-linked immunosorbent assay using monovalent and polyvalent antisera.

An indirect "sandwich" enzyme-linked immunosorbent assay (ELISA) using polyvalent and monovalent antisera was compared with the 50% complement fixation (CF50) test for the detection of foot-and-mouth disease (FMD) O, A, and C virus types. ELISA was more sensitive than CF50 tests when polyvalent antisera were used for detecting the 3 types of virus in epithelial samples, whereas ELISA using monovalent antisera was the least sensitive technique. The ELISA performed with polyvalent antisera was 9 times more sensitive for detecting FMD virus than that with monovalent antisera. However, viral isolation in cell culture was the most sensitive detection system. The combined use of ELISA with polyvalent antisera and cell culture inoculations was the most effective procedure for identifying FMD virus in epithelial samples from the field.     

Identification ofTomato Mottle Taino Begomovirus strains in Cuban potato fields

The presence of a begomovirus in potato plants with yellow mottle symptoms was determined for the first time in Cuba. The incidence of typical begomovirus-like symptoms in potato plants in some regions of Havana province (Güira de Melena, San José de las Lajas, Güines and Boyeros) during the growing seasons from 1992 to 1998 was in general low. However, in some cultivars belonging to the National Program for Potato Genetic Improvement, the incidence reached 100%. Yield losses, determined in 1992 and 1994, ranged as high as 19% to 56.33% depending on the cultivar. Characterization of the causal agent was done by light microscopy, host range (graft and mechanical transmission), DNA hybridizations, polymerase chain reaction, and restriction fragment length polymorphism analysis. Nucleotide sequence of the amplified fragments revealed the presence ofTomato mottle Taino virus. The virus was transmittedvia tubers and has been detected in mixed infections withPotato virus X and withPotato leaf roll virus. http://www.phytoparasitica.org posting Oct. 20, 2003. The first two authors contributed equally to this work.     

The organophosphorous insecticide chlorpyrifos affects the neuroepithelial junction, the bioelectric parameters of the skin of the frog Caudiverbera caudiverbera, and the structure of model cell membranes

The current study examines the acute effects of the organophosphorus pesticide chlorpyrifos on a sympathetic synapse of the frog Caudiverbera caudiverbera. Nerve stimulation was followed immediately by a transient increase in the short-circuit current (SCC) and in the potential difference (PD), which consisted of a rapid and then a slow component. Chlorpyrifos concentrations from 5 μM to 1.0 mM caused a dose-dependent block of both components to a 10% of their control values, which was reversed by washout. The pesticide blockade did not affect the skin response to noradrenaline. X-ray diffraction and fluorescence spectroscopy studies on membrane models showed marked phospholipid perturbation, which favors changes in ion channel conformation and interferes with receptor proteins, thus altering noradrenergic transmission and Cl⁻ secretion in the mucous glands. The foregoing results may be interpreted as a reversible inhibition of the neuroepithelial synapse to nerve stimulation, possibly due to non-specific lip μd-protein perturbation, interference with synaptic transmission, and transient Cl⁻ channel inactivation.     

Diagnosis and treatment of presumptive pyelonephritis in an Asian elephant (Elephas maximus).

A 37-yr-old female Asian elephant (Elephas maximus) presented with anorexia, restlessness, and dark-colored urine. Urinalyses showed hematuria, leukocyturia, isosthenuria, proteinuria, granular casts, and no calcium oxalate crystals. Bloodwork revealed azotemia. Urine culture revealed a pure growth of Streptococcus zooepidemicus resistant to sulfamethoxazole-trimethoprim but susceptible to cephalosporins. A presumptive diagnosis of pyelonephritis was made based on bloodwork, urinalysis, and urine culture. The animal was treated with intravenous ceftiofur, and intravenous and per rectum fluids were given for hydration. The elephant's attitude and appetite returned to normal, the abnormal blood parameters resolved, and urinary calcium oxalate crystals reappeared after treatment, supporting presumptive diagnosis. Follow-up ultrasonography revealed an abnormal outline of both kidneys with parenchymal hyperechogenicity and multiple uterine leiomyomas.     

Doxycycline treatment for <Emphasis Type="Italic">Dirofilaria immitis</Emphasis> in dogs: impact on <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and <Emphasis Type="Italic">Enterococcus</Emphasis> antimicrobial resistance

Doxycycline is an antibiotic that, in addition to the classic antibacterial use, is also prescribed to fight parasitic diseases, like heartworm disease in dogs. Despite the concern that the overuse of this antibiotic may decrease susceptibility of clinically important bacteria, the consequences of the prolonged doxycycline therapy in heartworm-infected dogs have never been studied before. We have analyzed the impact of this therapy on Staphylococcus aureus and Enterococcus antimicrobial resistance. In this study, 17 heartworm-infected dogs (10 that had completed the doxycycline treatment and 7 dogs that had not yet begun) were included. Twenty-four isolates of Staphylococcus aureus were obtained from two locations of each dog. After treatment, 73.3% of isolates were resistant to at least one antibiotic but only 22.2% of isolates before treatment. Most of doxycycline resistant isolates were obtained from dogs that have received treatment. Erythromycin resistance or intermediate susceptibility was detected in 45.6% of isolates, most of them from dogs after treatment. For Enterococci, 48 isolates were obtained from fecal samples (25 before treatment and 23 after treatment). Before treatment, 32% of isolates were resistant at least to one antibiotic while after, this data increase up to 65%. Comparing isolates before and after treatment, a clear increase in resistance to doxycycline (12% against 21.74%) and erythromycin (20% against 39.13%) was observed. Although the present work is a preliminary research, the results encourages the development of further studies to determinate the effect of prolonged doxycycline therapy on antimicrobial resistance.     

<Emphasis Type="SmallCaps">d</Emphasis>-Cloprostenol enhances estrus synchronization in tropical hair sheep

To compare the effects of PGF2α (dinoprost tromethamine) and d-cloprostenol in a two-dose protocol for estrus synchronization in hair sheep during breeding season in Yucatán, México, two experiments were conducted. In experiment 1, 61 cyclic hair sheep were divided into two groups: G1 (control n?=?30), two doses of 50 μg of dinoprost tromethamine IM with 12 days between applications, and G2 (n?=?31), two doses of 50 μg of d-cloprostenol IM at the same time interval. For determination of progesterone levels, 16 ewes from each group were randomly selected. In experiment 2, 70 cyclic hair sheep were assigned at the same treatments (G1 and G2, n?=?35) and 48 h after the second application, the ewes in estrus were detected by two vasectomized rams. Sheep with detected estrus were inseminated, and 45 days after, pregnant animals were identified by ultrasonography. An exact Fisher’s test was performed for the analysis of ewes in estrus (experiments 1 and 2) and number of pregnant ewes (experiment 2); for the comparison of time between end of treatment-estrus presentation, a survival analysis was used. Duration of estrus in hours was analyzed using a generalized mixed model (GLM) ANOVA whereas plasma progesterone concentrations were analyzed by non-linear regression. There were significant differences (P?<?0.05) in the proportion of ewes in estrus upon treatments (G1, 57% vs G2, 87% and G1, 37.1% vs G2, 65.7% in experiments 1 and 2, respectively), and between the end of treatment-onset estrus interval (P?<?0.01), survival curves showed the highest number of sheep in estrus between 40 and 48 h (G1, 43.7?+?8.05 h vs G2, 42.9?+?6.7 h, experiment 1). There were no significant differences (P?>?0.05) in duration of estrus (G1, 42?+?6.1 h, vs G2, 41.1?+?11.2 h, experiment 1) and pregnancy in the ewes that presented estrus, and were inseminated (G1, 38.4% vs 52.1%, experiment 2). With regard to concentrations of progesterone, significant differences (P?<?0.01) were found between treatments, and progesterone levels before the second application of d-cloprostenol were higher. In consideration of the results, it can be concluded that in a two-dose protocol of a luteolytic agent, more ewes presented estrus in response to d-cloprostenol compared to dinoprost tromethamine with similar pregnancy rates.