Determination of in vitro digestibility of forage species used in ruminant feeding

Within the evaluation of the quality of forage resources, the main parameter that defines it is the digestibility of dry matter, which together with the amount of neutral and acidic detergent fibers and crude protein constitutes the basic information to assess forages which are supplied in the diet of the cattle. This research was carried out at the University of Los Llanos (Villavicencio, Colombia), and its objective was to determine the digestibility of three forages in cattle through three different in vitro techniques: inoculation with ruminal fluid and with feces and enzymatic digestibility technique, making the comparison with the in situ technique in order to validate the techniques and equipment that are being used for these procedures. The following species were evaluated: Pennisetum purpureum (PP), Tithonia diversifolia (TD), and Bauhinia variegata (BV), assessing the curve and rate of degradation of dry matter (DM), neutral detergent fiber (NDF), and total protein (TP) (0 to 72 h). A design of repeated measures was used, under which the analysis of variance was carried out to determine the ranges of deviation between the techniques and thus establish the trend of the data; the variables evaluated were the DM, NDF, and TP digestibilities of the three forages using the four techniques (three in vitro and one in situ). After verifying the differences between the variances of the digestibilities and checking the sphericity assumption with the Mauchly test, multiple comparisons were made with the Bonferroni test with a significance of 5%. The digestibility of DM, NDF, and TP varied between 38.62 and 44.22, 54.18 and 66.97, and 47.54 and 57.05%; 49.07 and 70.70, 72.52 and 75.44, and 62.61 and 74.02%; 29.93 and 34.84, 26.21 and 70.88, and 25.67 and 50.60% respectively in forages PP, TD, and BV, depending on the technique used for their estimation. Despite finding statistically significant differences between several of the comparisons made in the digestibility techniques, a high coefficient of determination and a high correlation between the in vitro estimations with respect to the in situ estimation were found; therefore, it is possible to use these techniques routinely thus avoiding the need to have cattle with fistulae to perform digestibility tests, with enzymatic digestibility technique being the most practical one.

     

Effect of food on the pharmacokinetics of oral cefuroxime axetil in dogs

Cefuroxime axetil pharmacokinetic profile was investigated in 12 Beagle dogs after single intravenous and oral administration of tablets or suspension at a dose of 20 mg/kg, under both fasting and fed conditions. A three-period, three-treatment crossover study (IV, PO under fasting and fed condition) was applied. Blood samples were withdrawn at predetermined times over a 12-hr period. Cefuroxime plasma concentrations were determined by HPLC. Data were analyzed by compartmental analysis. No statistically significant differences were observed between formulations and feeding conditions on PK parameters. Independently of the feeding condition, absorption of cefuroxime axetil after tablet administration was low and erratic. The drug has been quantified in plasma in 3 out of 6 and 5 out of 6 dogs in the fasted and fed groups. For this formulation, the bioavailability (F), peak plasma concentration (C_max), and area under the concentration–time curve (AUC) of cefuroxime axetil were significantly enhanced (p < .05) by the concomitant ingestion of food (32.97 ± 13.47–14.08 ± 7.79%, 6.30 ± 2.62–2.74 ± 0.66 µg/ml, and 15.75 ± 3.98–7.82 ± 2.76 µg.hr/ml for F, C_max, and AUC in fed and fasted dogs, respectively), while for cefuroxime axetil suspension, feeding conditions affected only the rate of absorption, as reflected by the significantly shorter absorption half-life (T_½(a)) and time to peak concentration (T_max) (0.55 ± 0.27–1.15 ± 0.19 hr and 1.21 ± 0.22–1.70 ± 0.30 for T_½(a) and T_max in fed and fasted dogs, respectively). For cefuroxime axetil tablets, T > MIC (≤1 µg/ml) was <2 hr in fasted and ≈4 hr in fed animals, and for cefuroxime axetil suspension, T > MIC (≤1 µg/ml) was ≈5 hr and for T >MIC (≤4 µg/ml) was ≈2.5 hr for fasted and fed dogs, respectively. Cefuroxime axetil as a suspension formulation seems to be a better option than tablets. However, its short permanence in plasma could reduce its clinical usefulness in dogs.     

Pharmacokinetics of ciprofloxacin in sheep after single intravenous or intramuscular administration

Summary

The disposition and urinary excretion of ciprofloxacin (CIP) following intravenous (IV) or intramuscular (IM) administration of 7.5 mg/kg body weight in sheep (n = 5) was studied. The intravenous plasma concentration curve was best described pharmacokinetically by a two‐compartment open model, while the intramuscular administration data fitted better to a one‐compartment open model. Mean elimination half‐lives after IV and IM administration were 72 and 184 minutes, respectively. The absorption of intramuscularly administered CIP in sheep was fast: maximal plasma concentration (Cmax) was reached quickly (tmax 31.93 min) and attained values of 0.69 ± 0.27 mg/l. The bioavailability was 49%. The urinary data showed a significant decrease in the elimination rate constant of CIP when CIP was administered intramuscularly. The other parameters calculated did not display differences between the two routes of administration. The results obtained suggest that when CIP was administered by the IM route in the assayed dose, it was able to maintain serum concentrations above the MIC of most common pathogens over an 8‐hour period.     

Identification of helminth species by means of disc electrophoresis

Soluble proteins of 25 helminth species of the classes Trematoda, Cestoidea and Nematoda, were separated by disc electrophoresis using polyacrylamide gel columns. Differences between the species were investigated on the basis of Rm values of the bands. Protein spectra were complemented by the detection of lipoproteins and glycoproteins and by identification of LDH, SHD, peroxidase, esterase and alkaline phosphatase. On the basis of comparison of protein spectra of parasitic worms belonging to three taxonomic classes it was found by means of numerical taxonomy that individual classes are characterized by a certain number of proteins of the same migration properties.     

Penetration of ovicidal fungus Verticillium chlamydosporium through the Ascaris lumbricoides egg-shells

The course of penetration of the ovicidal fungus Verticillium chlamydosporium Goddard through the Ascaris lumbricoides egg-shells was studied in electron miorographs. The contact area between the egg and the fungus is smooth at the stage of the contact of the ovicidal fungus with the egg surface, as well as at the stage of adhesion. No special attaching organs are formed which indicates that no mechanical penetration of the fungal hypha through the egg-shells is involved. Traces of enzymatical activity of the fungus penetrating upright through the egg-shells can be observed during the penetration stage proper. At the consumption stage, the branching of the ovicidal fungus becomes more dense and some hyphae grow through the egg envelopes parallelly under the egg surface. The fungus grows through all parts of the infected egg, even through its contents.     

Identification and Characterisation of Bacteria Causing Soft-rot in Agave tequilana

Agave tequilana is the raw material for the production of the alcoholic beverage tequila. A bacterial disease has affected the A. tequilana crop in recent years. Previous reports based on colony and cell morphology, Gram stain and potato rot indicated that Erwinia sp. is the main pathogen. We isolated a several bacterial isolates capable of producing soft-rot symptoms in greenhouse pathogenicity assays. An extensive characterisation involving pathogenicity tests, fatty acid profile, metabolic and physiological properties, ribosomal DNA sequence and intergenic transcribed spacer amplification (ITS-PCR) and restriction banding pattern (ITS-RFLP) was made of each isolate. Three different species: Erwinia cacticida, Pantoea agglomerans and Pseudomonas sp. were identified. Fatty acid and metabolic profiles gave low similarity values of identification but 16S rDNA sequence, ITS-PCR and ITS-RFLP confirmed the identification of E. cacticida. In the phylogenetic tree, E. cacticida from blue agave was grouped neither with E. cacticida type strains nor with Erwinia carotovora. This is the first report that associates E. cacticida with A. tequilana soft-rot symptoms.     

Subolesin/akirin orthologs from Ornithodoros spp. soft ticks: cloning, RNAi gene silencing and protective effect of the recombinant proteins

Subolesin/akirin is a well characterized protective antigen highly conserved across vector species and thus potentially useful for the development of a broad-spectrum vaccine for the control of arthropod infestations including hard ticks, mosquitoes, sand flies and the poultry red mite Dermanyssus gallinae. Soft ticks could be also targeted by this vaccine if proved that the soft tick subolesin orthologs are conserved and induce protective immune responses too. However, to date no soft tick subolesin orthologs have been fully characterized nor tested as recombinant antigens in vaccination trials. The objectives of the present work were to clone and characterize the subolesin orthologs from two important vector species of soft ticks as Ornithodoros erraticus and O. moubata, to evaluate the effect of subolesin gene silencing by RNAi, and to test the protective value of the recombinant antigens in vaccination trials. The obtained results demonstrate that both soft tick subolesins are highly conserved showing more than 69% and 74% identity with those of hard ticks in their nucleotide and amino acid sequences, respectively. Additionally, we demonstrate that both soft ticks possess fully operative RNAi machinery, and that subolesin gene silencing by dsRNA injection inhibits oviposition indicating the involvement of subolesin in tick reproduction. Finally, vaccination with the recombinant soft tick subolesins induced a partial protective effect resulting in the reduction of the oviposition rate. These preliminary results encourage further studies on the use of recombinant subolesins as vaccines for the control of soft tick infestations, either alone or in combination with other specific molecules.     

Effects of the Administration of Lactobacilli,Maltodextrins and Fructooligosaccharides upon the Adhesion of <Emphasis Type="Italic">E. coli</Emphasis> O8:K88 to the Intestinal Mucosa and Organic Acid Levels in the Gut Contents of Piglets

The influence of the administration of Lactobacillus plantarum, maltodextrin Maldex 150 and Raftifeed IPX fructooligosaccharides on the inhibition of adhesion of E. coli O8:K88 to the mucosa of the jejunum, ileum and colon as well as on the organic acid levels was investigated in 33 conventional piglets. The counts of E. coli K88 adhering to the jejunal mucosa were significantly decreased (p < 0.05) in Lact. plantarum + Maldex 150 and Lact. plantarum + Maldex 150 + Raftifeed IPX groups. The counts of E. coli K88 adhering to the colonic mucosa of Lact. plantarum + Maldex 150 + Raftifeed IPX and Lact. plantarum + Raftifeed IPX groups were significantly lower (p < 0.05) than in Lact. plantarum and Lact. plantarum + Maldex 150 animals. The acetic acid levels in the ileum and colon of the Lact. plantarum + Maldex 150 + Raftifeed IPX group and Lact. plantarum + Raftifeed IPX group were significantly higher (p < 0.05) than in the Lact. plantarum and Lact. plantarum + Maldex 150 group. The combination of Lact. plantarum, maltodextrin Maldex 150 and Raftifeed IPX proved to be the most effective one to inhibit the counts of E. coli O8:K88 adhering to the intestinal mucosa of the jejunum and colon of conventional piglets.