Ovum pick-up and in vitro maturation in spotted paca (Cuniculus paca—Linnaeus, 1766)

We tested FSHp, eCG and FSHp + eCG to establish ovum pick-up (OPU) and in vitro maturation method in spotted paca. Eight healthy adult females were subjected to each of four treatments to stimulate ovarian follicular growth. All females were subjected to a hormonal protocol using a single dose of 45 mg of injectable progesterone and single intramuscular injection of 0.075 mg d-cloprostenol on day 6. Ovarian stimulation was carried out as follows: in Group TFE (FSHp and eCG), animals were treated with a single dose of 80 mg of FSHp and 200 IU of eCG intramuscularly on day 6 after the application of progesterone; in Group TF (FSHp), they were treated with a single dose of 80 mg of FSHp intramuscularly on day 6 after application of progesterone; in Group treatment eCG, they were treated with 200 IU of eCG intramuscularly on day 6 after application of progesterone; and in Group TC (saline solution), 1 ml of saline solution was administered to control does. The OPU was performed between 22 and 26 hr after gonadotropin treatments. All recovered oocytes were placed into maturation media and incubated for 24 hr. There were no differences among the mean number of observed follicles, aspirated follicles and oocytes recovered per treatment. Oocyte maturation rates did not differ among groups, except, TF and treatment eCG oocytes had greater maturation rates than TC oocytes. In this study, gonadotropin administration failed to superovulate treated does and increase oocyte retrieval efficiency. Despite the feasibility of the procedure, further studies are needed to develop and refine hormonal protocols for oocyte recovery and in vitro maturation in this species.     

A new type of pterocarpanquinone that affects Toxoplasma gondii tachyzoites in vitro

Toxoplasma gondii, the agent of Toxoplasmosis, is an obligate intracellular protozoan able to infect a wide range of vertebrate cells, including nonprofessional and professional phagocytes. Therefore, drugs must have intracellular activities in order to control this parasite. The most common therapy for Toxoplasmosis is the combination of sulfadiazine and pyrimethamine. This treatment is associated with adverse reactions, thus, the development of new drugs is necessary. In previous studies, naphthoquinone derivatives showed anti-cancer activity functioning as agents capable of acting on groups of DNA, preventing cancer cells duplication. These derivatives also display anti-parasitic activity against Plasmodium falciparum and Leishmania amazonensis. The derivative pterocarpanquinone tested in this work resulted from the molecular hybridization between pterocarpans and naphtoquinone that presents anti-tumoral and anti-parasitic activities of lapachol. The aim of this work was to determine if this derivative is able to change T. gondii growth within LLC-MK2 cells. The drug did not arrest host cell growth, but was able to decrease the infection index of T. gondii with an IC(50) of 2.5 μM. Scanning and transmission electron microscopy analysis showed morphological changes of parasites including membrane damage. The parasite that survived tended to encyst as seen by Dolichos biflorus lectin staining and Bag-1 expression. These results suggest that pterocarpanquinones are drugs potentially important for the killing and encystment of T. gondii.     

Molecular characterization of the VP1 gene of a Mexican isolate of infectious pancreatic necrosis virus

The presence of infectious pancreatic necrosis virus (IPNV) in salmonids predominantly produces a high mortality rate in first-feeding fry. Genomic analysis of the vp2 gene sequence is most commonly used to determine the genetic diversity of IPNV isolates. Recently, information obtained from the vp1 gene allowed for efficient analysis of the genetic diversity of IPNV. In this study, the vp1 gene from a Mexican IPNV isolate was characterized and compared with IPNV isolates from Europe, North America, and Asia. The results indicate that the Mexican isolate is most closely related genetically to the 2310 strain from Spain.     

High prevalence of F4<Superscript>+</Superscript> and F18<Superscript>+</Superscript><Emphasis Type="Italic">Escherichia coli</Emphasis> in Cuban piggeries as determined by serological survey

Little information is available on the prevalence of swine enteropathogens in Cuba where diarrheic diseases are responsible for 31% and 37% of the total mortality during the neonatal and postweaning periods. F4⁺ and F18⁺ enterotoxigenic Escherichia coli and F18⁺ verotoxigenic E. coli induce diarrhea and edematous disease in pigs, but their distribution has never been thoroughly studied in the Cuban swine population. Therefore, the present study estimated the prevalence of F4- and F18-specific antibodies in sera of 1,044 6-month-old gilts distributed in 34 piggeries spread over the Cuban territory. For the data analysis, which included the optical density of individual samples tested by ELISA, random-effects models and a mixture model in R (package “mixAK”; Komárek, Computational Statistics and Data Analysis 53:3932–3947, 2009) were fitted. Low, moderate, and high levels of F4-specific antibodies were found in 67.6%, 26.8%, and 5.6% of the gilts, while 66.4% and 33.6% of gilts showed low and high levels of F18-specific antibodies. Hereby, we show that F4⁺ and F18⁺ E. coli are highly prevalent as potential enteropathogens in Cuban piggeries.     

Applications of a cell culture system for studying the interaction of Anaplasma marginale with tick cells

A cell culture system for the tick-borne rickettsia Anaplasma marginale offers new opportunities for research on this economically important pathogen of cattle. A. marginale multiplies in membrane-bound inclusions in host cells. Whereas erythrocytes appear to be the only site of infection in cattle, A. marginale undergoes a complex developmental cycle in ticks and transmission occurs via the salivary glands during feeding. We recently developed a cell culture system for A. marginale using a cell line derived from embryos of Ixodes scapularis. Here we review the use of this cell culture system for studying the interaction of A. marginale with tick cells. Several assays were developed using the A. marginale/tick cell system. An adhesion assay was developed for the identification of proteins required by A. marginale for adhesion to tick cells. The effect of antibodies against selected major surface proteins in inhibiting A. marginale infection was tested in an assay that allowed further confirmation of the role of surface proteins in the infection of tick cells. A drug screening assay for A. marginale was developed and provides a method of initial drug selection without the use of cattle. The culture system was used to test for enhancing effects of tick saliva and saliva components on A. marginale infection. The tick cell culture system has proved to be a good model for studying A. marginale-tick interactions. Information gained from these studies may be applicable to other closely related tick-borne pathogens that have been propagated in the same tick cell line.     

In vitro activity of the nonsteroidal anti-inflammatory drug indomethacin on a scuticociliate parasite of farmed turbot

The scuticociliatosis produced by the endoparasite Philasterides dicentarchi is a severe parasitic infection of farmed turbot (Scophthalmus maximus) characterized by several histopathological effects including extensive inflammation. Indomethacin is a nonsteroidal anti-inflammatory drug that specifically inhibits synthesis of the proinflammatory mediator prostaglandins. The effect of indomethacin on the in vitro growth of P. dicentrarchi was investigated. In vitro growth of the scuticociliate was significantly inhibited by treatment with 100 microM indomethacin for 48 h. Higher concentrations of indomethacin (mM levels) did not affect the gelatinolytic activity of the cysteine proteinases of P. dicentrarchi. In vitro treatment with 25, 50 or 100 microM indomethacin for 3 days did not significantly affect the enzymatic activity of cysteine proteinases, as assayed with p-nitroanilide as substrate. Immunoblot analysis with anti-cysteine proteinase antibodies revealed an increase in proteinase expression (molecular weights of 80, 32 and 40-45 kDa) in parasite lysates originating from in vitro cultures incubated with 25 microM indomethacin for 72 h. Degradation of genomic DNA of the ciliates was observed in cultures incubated with 100 microM indomethacin for 1, 3 and 7 days. The results suggest that indomethacin is capable of inhibiting in vitro growth of the scuticociliate P. dicentrarchi by a mechanism related to the induction of programmed cell death, without affecting the enzymatic activation of parasite proteinases, which demonstrates the potential therapeutic use of this drug in the control of turbot scuticociliatosis.     

Neurological disorder in dairy cattle associated with consumption of beer residues contaminated with Aspergillus clavatus.

A neurological syndrome in dairy cattle associated with consumption of moldy beer residues is described. The disease occurred on 1 farm in late June 2001, during winter. Six heifers and 1 cow out of 45 cattle were affected during a 3-week period. The affected animals died spontaneously or were euthanized approximately 2-14 days after the onset of clinical signs. The clinical signs were characterized by flaccid paralysis and gait abnormalities. Clinical signs were more pronounced after exercise and included stiff and unsteady gait, knuckling at the fetlocks of the hind limbs, frequent falling, inability to rise, muscular tremors, especially of the head and the hindquarters, and drooling. Main necropsy findings included degenerative and necrotic changes of the larger medial muscle groups of the hindquarters, i.e., adductor, pectineus, quadriceps femoris, rectus femuris, sartorius, semimembranosus, semitendinosus, and vastus medialis, and of the forequarters, including pectoralis descendens, pectoralis ascendens, and transversus pectoralis. The main histologic findings consisted of degenerative and necrotic neuronal changes (chromatolysis) of varying severity and extent affecting selected nuclei of the brainstem and neurons of the ventral horns of the spinal cord. Similar microscopic lesions were observed in the neurons of the spinal cord of 1 experimental sheep force-fed for 35 days with 1 kg/day of the same batch of foodstuff that was originally fed to the cattle. Coarse white or gray lumps, interpreted as mycelia, were observed in the beer by-product. Aspergillus clavatus was the dominant fungus isolated. Deaths ceased after the consumption of beer residue was discontinued. Recovery from illness was observed in 1 animal. The diagnosis was based on epidemiological data, clinical signs, necropsy findings, histological lesions, dosing trial, and mycology. A similar condition caused by consumption of barley by-products, sprouted wheat, corn sprouts, and beetroot screenings contaminated with A. clavatus has been reported in cattle and sheep worldwide.     

Effect of first-pass hepatic metabolism on the disposition of levamisole after intravenous administration in rabbits

OBJECTIVE: To evaluate the contribution of first-pass hepatic metabolism of levamisole on levamisole disposition in rabbits. ANIMALS: 30 male New Zealand White rabbits. PROCEDURES: Rabbits were randomly placed into 2 groups. Rabbits in the first group received levamisole via the marginal ear vein at the following 3 doses: 12.5, 16, and 20 mg/kg (5 rabbits for each dose). Rabbits of the second group received levamisole via the jejunal vein at the same doses (5 rabbits each). During the following 240-minute period, plasma samples were obtained and quantified for levamisole concentrations by reversed-phase high-performance liquid chromatography. RESULTS: No significant differences were found between pharmacokinetic parameters calculated by compartmental or noncompartmental analysis. Mean hepatic extraction ratio ranged from -0.044 to 0.017 and from 0.020 to 0.081 when area under the plasma concentration-time curve values were obtained after compartmental or noncompartmental analysis, respectively. After compartmental analysis, plasma concentration decreased bi-exponentially. Mean pharmacokinetic parameter values were as follows for each dose (12.5, 16, and 20 mg/kg, respectively): after levamisole administration via the marginal ear vein, volume of distribution at steady state (Vss) = 4.26, 4.33, and 3.20 L/kg; total body clearance (CI) = 49.04, 43.77, and 39.26 mL/kg x min; and half-life associated with beta-phase (t1/2beta) = 77.93, 85.39, and 69.79 minutes. After levamisole administration via the jejunal vein, Vss = 4.38, 2.85, and 2.97 L/kg; CI = 48.14, 42.40, and 39.69 mL/kg x min; and t1/2b = 101.9, 76.71, and 76.13 minutes. CONCLUSIONS: Levamisole has a low degree of hepatic extraction in rabbits.     

Carbon Stocks and Fluxes in Rangelands of the Río de la Plata Basin

Grasslands are one of the most modified biomes on Earth. Land use changes had a large impact on carbon (C) stocks of grasslands. Understanding the impact of land use/land cover changes on C stocks and fluxes is critical to evaluate the potential of rangeland ecosystem as C sinks. In this article we analyze C stocks and fluxes across the environmental gradients of one of the most extensive temperate rangeland areas: the Río de la Plata Grasslands (RPG) in South America. The analysis summarizes information provided by field studies, remote sensing estimates, and modeling exercises. Average estimates of aboveground net primary production (ANPP) ranged from 240 to 316 g C· m^?2·yr^?1. Estimates of belowground NPP (BNPP) were more variable than ANPP and ranged from 264 to 568 g C· m^?2·yr^?1. Total Carbon ranged from 5 004 to 15 008 g C· m^?2. Plant biomass contribution to Total Carbon averaged 13% and varied from 9.5% to 27% among sites. The largest plant C stock corresponded to belowground biomass. Aboveground green biomass represented less than 7% of the plant C. Soil organic carbon (SOC) was concentrated in the slow and passive compartments of the organic matter. Active soil pool represented only 6.7% of the SOC. The understanding of C dynamics and stocks in the RPG grasslands is still partial and incomplete. Field estimates of ANPP and BNPP are scarce, and they are not based on a common measurement protocol. Remotely sensed techniques have the potential to generate a coherent and spatially explicit database on ANPP. However, more work is needed to improve estimates of the spatial and temporal variability of radiation use efficiency. The absence of a flux tower network restricts the ability to track seasonal changes in C uptake and to understand fine-scale controls of C dynamics.