Retrospective study on the occurrence of the feline lungworms Aelurostrongylus abstrusus and Troglostrongylus spp. in endemic areas of Italy

Aelurostrongylus abstrusus is a metastrongyloid nematode infesting the respiratory system of domestic cats worldwide. Troglostrongylus brevior and Troglostrongylus subcrenatus, two lungworms thought to infest wild felids, have been found recently in domestic cats from Spain and Italy. These unexpected findings have raised doubts about the assumed past and present occurrence of Troglostrongylus spp., especially T. brevior, in domestic hosts and suggest that there may have been missed detection or misdiagnosis. The present retrospective study evaluated the presence of lungworms in cats from Italy with a diagnosis of respiratory parasitism or with compatible lung lesions from 2002 to 2013. Sixty-eight samples of DNA and larvae from cats with a diagnosis of aelurostrongylosis, and 53 formalin-fixed paraffin-embedded lung samples from cats confirmed as lungworm infested or with compatible lesions, were investigated using two DNA-based assays specific for A. abstrusus or T. brevior. All DNA and larval samples were positive for A. abstrusus and one was additionally positive for T. brevior. Most paraffin-embedded lung tissues were positive only for A. abstrusus, but two samples tested positive for both lungworms and one for T. brevior only. This study supports the major role of A. abstrusus in causing feline respiratory parasitism in endemic areas of Italy.     

Pharmacokinetics,plasma protein binding and bioavailability of moxifloxacin in Muscovy ducks after different routes of administration

In this study the disposition kinetics and plasma availability of moxifloxacin in Muscovy ducks after single intravenous (i.v.), intramuscular (i.m.) and oral (p.o.) administrations of 5 mg kg^?1 b.wt. were investigated. The concentrations of moxifloxacin in the plasma were measured using high-performance liquid chromatography (HPLC) with fluorescence detection on samples collected at frequent intervals after drug administration. Following intravenous injection, the decline in plasma drug concentration was bi-exponential with half-lives of (t_1/2α) 0.22 ± 0.10 h and (t_1/2β) 2.49 ± 0.26 h for distribution and elimination phases, respectively. The volume of distribution at steady-state (V_dss) was 1.02 ± 0.14 l kg^?1 and the total body clearance (Cl_tot) was 0.32 ± 0.11 l kg^?1 h^?1, respectively. After intramuscular and oral administration of moxifloxacin at the same dose the peak plasma concentrations (C_max) were 2.38 ± 0.43 and 2.11 ± 0.36 μg ml^?1 and were obtained at 1.47 ± 0.26 and 1.83 ± 0.16 h (T_max), respectively, the elimination half-lives (T_1/2el) were 3.14 ± 0.42 and 2.63 ± 0.44 h, respectively, and AUC_0–24 were 15.87 ± 2.35 and 14.52 ± 2.37 μg ml^?1 h^?1, respectively. The systemic bioavailabilities were 96.36 ± 11.54% and 86.79 ± 12.64%, respectively. In vitro plasma protein binding percent was 32%. We concluded that moxifloxacin might be clinically interesting alternative for the treatment of most sensitive bacterial infections in Muscovy ducks.     

Isolation of active compounds from methanol extracts of Toddalia asiatica against Ichthyophthirius multifiliis in goldfish (Carassius auratus)

The parasitic ciliate Ichthyophthirius multifiliis infests all species of freshwater fish and can cause severe economic losses in fish breeding. The present study aims to evaluate the antiparasitic activity of the active components from Toddalia asiatica against I. multifiliis. Bioassay-guided fractionation and isolation of compounds with antiparasitic activity were performed on the methanol extract of T. asiatica yielding two bioactive compounds: chelerythrine and chloroxylonine identified by comparing spectral data (NMR and ESI-MS) with literature values. Results from in vitro antiparasitic assays revealed that chelerythrine and chloroxylonine could be 100% effective against I. multifiliis at the concentration of 1.2 mg L^?1 and 3.5 mg L^?1, with the median effective concentration (EC₅₀) values of 0.55 mg L^?1 and 1.90 mg L^?1 respectively. In vivo experiments demonstrated that fish treated with chelerythrine and chloroxylonine at the concentrations of 1.8 and 8.0 mg L^?1 carried significantly fewer parasites than the control (P < 0.05). The acute toxicity (LC₅₀) of chelerythrine for goldfish was 3.3 mg L^?1.     

The use of eugenol against Aeromonas hydrophila and its effect on hematological and immunological parameters in silver catfish (Rhamdia quelen)

The aim of this study was to evaluate the activity of eugenol against the fish pathogen Aeromonas hydrophila and eugenol's effect on hematological and natural immune parameters in silver catfish (Rhamdia quelen). In vitro, eugenol showed weak activity against A. hydrophila, but in vivo, at a subinhibitory concentration (10 mg L^?1), it promoted survival in infected silver catfish. Eugenol (50 μg mL^?1) reduced the hemolytic activity of A. hydrophila supernatant in vitro in fish erythrocytes. Subjecting catfish to eugenol baths (5 and 10 mg L^?1) for five days did not alter the hematological and immunological parameters studied in this work. Based on these results, eugenol can be used to treat or prevent bacterial diseases in fish.     

Supplementing Guinea grass with fresh sweet potato foliage for milk production by Bunaji and N'Dama cows in early lactation

Forage from three sweet potato cultivars (A = TIS-87/0087; B = TIS-8164; C = TIS-2532.OP.1.13 at 30% daily dry matter intake), dried brewers' grains (DBG) and cottonseed meal (CSM) each at 2.5 kg were supplemented to Guinea grass (GG) to form four diets: Diet A = GG + TIS-87/0087; Diet B = GG + TIS-8164; Diet C = GG + TIS-2532.OP.1.13, and Diet D = GG + DBG + CSM (as control). Treatments were assigned as 4 × 4 Latin squares design over 60 days (10-day adaptation and 5-day sampling) using Bunaji and N'Dama cows in early lactation. The 48-h rumen dry matter (DM) degradation ranged (P < 0.01) from 407 g kg^? 1 DM for GG to 791 g kg^? 1 DM for sweet potato cultivar TIS-87/0087. Bunaji dry matter intake varied (P < 0.05) between 7.1 kg day^? 1 in Diet B and 8.9 kg day^? 1 in Diet D, but was similar (P > 0.05) among diets for the N'Dama cows. The metabolisable energy (ME) intakes were higher for Diet D although, it recorded the least efficiency of ME utilization for milk production. Milk yields were significantly (P < 0.01) higher in the Bunaji than the N'Dama cows, which is typical of their true breed differences. Total solids, ash, protein, fat, and sugar contents of the milk were similar among diets for both cow breeds, except Bunaji ash contents that ranged (P < 0.05) from 0.77 g 100 g^? 1 for Diet B to 0.83 g 100 g^? 1 for Diet D. The results suggest that sweet potato forage could be utilized as whole or partial replacement for DBG and CSM to save cost under smallholder farming systems.     

Animal and Vegetation Response to Modified Intensive–Early Stocking on Shortgrass Rangeland

A comparison of animal gains and vegetation trends was made from 2002–2008 between a continuous season-long stocking (SLS) system and a modified intensive–early stocking system (IES) with late-season grazing (IES 1.6× + 1; 1.6 times the number of animals of the SLS system from May 1 to July 15, and 1 times the number of animals of SLS from July 15 to October 1) on shortgrass native rangeland of western Kansas. The continuous season-long stocked system placed animals at a density of 1.37 ha · steer^?1 from May through October, or 2.63 animal unit months (AUM) · ha^?1, whereas the intensive–early stocked system with late-season grazing (3.33 AUM · ha^?1) stocked pastures at 0.85 ha · steer^?1 from May through the middle of July, and then stocked pastures at 1.37 ha · steer^?1 for the remainder of the grazing season by removing the heaviest animals mid-July each yr. Average daily gains (0.78 vs. 0.70 kg · d^?1, P = 0.039) and total animal gain (58 vs. 52 kg, P = 0.042) were different between the continuous season-long stocked and the intensive–early stocked animals during the first half of the grazing season. No difference was found between average daily gain (0.61 vs. 0.62 kg · d^?1, P = 0.726) and total animal gain (48 vs. 49 kg, P = 0.711) for the continuous season-long stocked and intensive–early stocked with late-season grazing animals during the last half of the season. Total individual animal gain (106 vs. 101 kg, P = 0.154) and average daily gain (0.70 vs. 0.66 kg · d^?1, P = 0.152) was not different between the continuous season-long stocked and the intensive–early stocked system animals that were on pasture the entire grazing season. Total beef gain on a land-area basis (96 vs. 77 kg · ha^?1, P = 0.008) was greater for the modified intensive–early stocked system with late-season grazing with greater animal densities. Changes in residual biomass and most key vegetation components at the end of the grazing season were not different between the two systems.     

Cryptosporidium scrofarum n. sp. (Apicomplexa: Cryptosporidiidae) in domestic pigs (Sus scrofa)

We describe the morphological, biological, and molecular characteristics of Cryptosporidium pig genotype II and propose the species name Cryptosporidium scrofarum n. sp. to reflect its prevalence in adult pigs worldwide. Oocysts of C. scrofarum are morphologically indistinguishable from C. parvum, measuring 4.81–5.96 μm (mean = 5.16) × 4.23–5.29 μm (mean = 4.83) with a length to width ratio of 1.07 ± 0.06 (n = 400). Oocysts of C. scrofarum obtained from a naturally infected pig were infectious for 8-week-old pigs but not 4-week-old pigs. The prepatent period in 8-week-old Cryptosporidium-naive pigs was 4–6 days and the patent period was longer than 30 days. The infection intensity of C. scrofarum in pigs was generally low, in the range 250–4000 oocysts per gram of feces. Infected pigs showed no clinical signs of cryptosporidiosis and no pathology was detected. Cryptosporidium scrofarum was not infectious for adult SCID mice, adult BALB/c mice, Mongolian gerbils (Meriones unguiculatus), southern multimammate mice (Mastomys coucha), yellow-necked mice (Apodemus flavicollis), or guinea pigs (Cavia porcellus). Phylogenetic analyses based on small subunit rRNA, actin, and heat shock protein 70 gene sequences revealed that C. scrofarum is genetically distinct from all known Cryptosporidium species.     

Characterization of Mannheimia haemolytica biofilm formation in vitro

Mannheimia haemolytica is the primary bacterial agent in the bovine respiratory disease complex. It is thought that M. haemolytica colonizes the tonsillar crypts of cattle as a commensal and subsequently descends into the lungs to cause disease. Many bacterial species persist in the host as biofilms. There is limited information about the ability of M. haemolytica to form biofilms. The aim of this study was to develop an in vitro model for M. haemolytica biofilm formation. We found that M. haemolytica required at least 36 h to form robust biofilms on plastic in vitro when incubated in RPMI-1640 tissue culture medium at 37 °C, with maximal biofilm formation being evident at 48 h. Biofilm formation was inhibited by adding the monosaccharides d(+) galactose and d(+) mannose to the growth medium. Addition of antibodies to the M. haemolytica surface protein OmpA also reduced biofilm formation. Upon evaluating the macromolecules within the biofilm extracellular polymeric substance we found it contained 9.7 μg/cm² of protein, 0.81 μg/cm² of total carbohydrate, and 0.47 μg/cm² of extracellular DNA. Furthermore, proteinase K treatment significantly decreased biofilms (P < 0.05) while α-amylase and micrococcal nuclease decreased biofilms to a lesser extent. M. haemolytica biofilm cells were more resistant than planktonic cells to the antibiotics florfenicol, gentamicin, and tulathromycin. These results provide evidence that M. haemolytica can form biofilms, which could contribute to its ability to persist as a commensal in the bovine upper respiratory tract.     

Pharmacokinetic,metabolism and withdrawal time of orphenadrine in camels (Camelus dromedarius) after intravenous administration

The pharmacokinetics of orphenadrine (ORPH) following a single intravenous (i.v.) dose was investigated in six camels (Camelus dormedarius). Orphenadrine was extracted from the plasma using a simple sensitive liquid–liquid extraction method and determined by gas chromatography/mass spectrometry (GC/MS). Following i.v. administration plasma concentrations of ORPH decline bi-exponentially with distribution half-life (t_1/2α) of 0.50 ± 0.07 h, elimination half-life (t_1/2β) of 3.57 ± 0.55 h, area under the time concentration curve (AUC) of 1.03 ± 0.10 g/h l⁻¹. The volume of distribution at steady state (Vd_ss) 1.92 ± 0.22 l kg⁻¹, volume of the central compartment of the two compartment pharmacokinetic model (V_c) 0.87 ± 0.09 l kg⁻¹, and total body clearance (Cl_T) of 0.60 ± 0.09 l/h kg⁻¹. Three orphenadrine metabolites were identified in urine samples of camels. The first metabolite N-desmethyl-orphenadrine resulted from N-dealkylation of ORPH with molecular ion m/z 255. The second N,N-didesmethyl-orphenadrine, resulted from N-didesmethylation with molecular ion m/z 241. The third metabolite, hydroxyl-orphenadrine, resulted from the hydroxylation of ORPH with molecular ion m/z 285. ORPH and its metabolites in camel were extensively eliminated in conjugated form. ORPH remains detectable in camel urine for three days after i.v. administration of a single dose of 350 mg orphenadrine aspartate.     

Performance of spring-calving beef suckler cows and their progeny to slaughter on intensive and extensive grassland management systems

The performance of rotationally grazed beef suckler cows and their progeny to slaughter on two lowland grassland management systems differing in stocking rate (SR) and fertiliser nitrogen (N) level was compared over eight years. The two Systems were 1) Intensive (INT): SR of 0.56 (bull production) or 0.71 (steer production) ha cow^? 1 unit, 211 kg fertiliser N ha^? 1, two silage harvests, and 2) Extensive (EXT): SR of 0.69 (bull production) or 0.88 (steer production) ha cow^? 1 unit, 97 kg fertiliser N ha^? 1 and one staggered silage harvest. A cow unit was defined as a cow plus progeny to slaughter. On the silage harvesting area, the mean application rate for fertiliser N was 110 and 80 kg ha^? 1 for first and second harvests, respectively. Herbage dry matter digestibility both pre- and post-grazing was similar (P > 0.05) for the two systems, whereas herbage crude protein concentrations were generally significantly lower for the EXT than the INT system. There was no difference (P > 0.05) between the Systems in cow live weight, body condition score or their changes or in calf live weight gain from birth to weaning. Post-weaning, live weight gain, slaughter weight, carcass weight, kill-out proportion, estimated carcass gain, carcass conformation score or carcass fat score did not differ (P > 0.05) between the systems for heifer, steer or bull progeny. It can be concluded that similar animal performance levels can be expected in an extensive grassland-based suckler calf-to-beef system compatible with the EU, Rural Environmental Protection Scheme as that attained in a more intensive System comprising of both a moderately high SR (~ 1.25 higher) and fertiliser N application (~ 2.1 higher).     

Modelling the lactation curve of rabbit does: Towards a model including fit suitability and biological interpretation

This work proposes an adequate empirical model for the 28-day lactation curve of rabbit does, including fit suitability and biological interpretation. A total of 15,400 test-day milk records were used, corresponding to 550 lactations collected from 134 hybrid New Zealand × Californian rabbit does during five consecutive lactations. To develop this model, five different functions were compared (quadratic, potential, beta-modified, gamma and Gauss models), evaluating their fitting ability to mean and individual lactation curves, and the suitability of their parameters to gather the sources of variation (genetic selection level, type of diet, parity order and gestation overlapping degree) on lactation curve shape. The possible relationship between model parameters and main performance traits was also evaluated. From the results of the present work, it may be concluded that beta-modified equation [Milk yield (g/day) = k × (day/30)^a × (1 −  (day/30))^b] could be proposed as an alternative to quadratic models for daily milk yield prediction of reproductive rabbit does. When compared to quadratic models, beta-modified model give a slightly better fit to average (R² = 0.986 vs. 0.985; RMSE = 5.648 vs. 5.813) and individual (Residuals = 21.31 vs. 21.37 g; Mean square prediction error = 883.0 vs. 897.2 g²) lactation curves, especially of those curves showing a lower lactation peak height and a greater persistence of milk yield. However, the most important advantage of the beta-modified model was the greater biological interpretation of its parameters (k regulates the curve height, while a and b regulate the milk yield of ascending and descending period, respectively) and the ability to gather curve changes. This latter aspect is revealed by the relationship of the parameters with main performance traits of lactating does (energy intake, live weight and body reserves mobilisation). Although further research on developing an optimal model is needed, the use of this type of models could provide additional information for a better understanding of the curve shape effect on the performance, body condition and health of reproductive rabbit does.     

Trichinella spiralis—A potential anti-tumor agent

Murine forestomach carcinoma (cell line MFC), ascitic hepatoma (cell line H22) and sarcoma (cell line S180) solid tumor models were used to test the anti-tumor effect of Trichinella spiralis in vivo. Mice previously infected by oral administration of 400 viable T. spiralis larvae per mouse for 7 days were grafted with various solid tumor cell lines. Other groups of tumor-bearing mice were given caudal vein injection of crude extracts of adult and newborn larvae at 17.5, 35.0 or 70.0 mg kg^?1. These treatments to inhibit tumor growth were dose-dependent (p < 0.05). The anti-proliferative activity of crude T. spiralis extract was examined in vitro at 0.035, 0.070 or 0.140 mg ml^?1 using MFC, H22, S180, human chronic myeloid leukemia cell line (K562) and hepatoma cell line (H7402), tumor cell proliferation in vitro was measured by methyl thiazolium stain and was inhibited in dose-dependent manner (p < 0.05). At the same doses, crude T. spiralis extracts induced apoptosis of K562 and H7402 as detected by DNA fragmentation. Cell cycle analysis indicated that crude T. spiralis extracts, at 0.140 mg ml^?1, arrested the cell cycle of K562 and H7402 in G1 or S phase. It is concluded that T. spiralis contains anti-tumor active agent.     

Antinociceptive efficacy and plasma concentrations of transdermal buprenorphine in dogs

To assess the antinociceptive efficacy of transdermal (TD) buprenorphine (B) in dogs, a prospective, positive-controlled experimental study was performed in 10 healthy Beagles. In an open label crossover design, the dogs initially received intravenous B (IVB, 0.02 mg kg^?1) as a positive control, followed by TDB (52.5 μg h^?1) 4 months later. Blood was collected at regular intervals for determination of the plasma concentrations of B ([B]) and its metabolite norbuprenorphine. The antinociceptive efficacy was assessed using thermal and mechanical models of nociception. The peak concentration [B] was 1.54 ng mL^?1 (±1.98) 60 h after TDB application, although three dogs had no measurable [B] after TDB. Maximum thermal threshold (TT) was 52.6 °C (±0.48) at 1 h after IVB administration and 51.63 °C (±1.01) 72 h after TDB application. The significant increase in TT indicated that effective antinociception was achieved beyond 36 h after the application of TDB, lasting until patch removal. There was hysteresis between [B] and the antinociceptive effect.     

Immune responses of orange-spotted grouper,Epinephelus coioides,against virus-like particles of betanodavirus produced in Escherichia coli

Betanodaviruses are the causative agents of viral nervous necrosis (VNN), a serious disease of cultured marine fish worldwide. Virus-like particles (VLPs) are one of the good novel vaccine candidates to control this disease. Until now, betanodavirus vaccine studies mainly focused on the humoral immune response and mortality after virus challenge. However, little is known about the activation of genes responsible for cellular and innate immunity by vaccines. In the present study, VLPs of orange-spotted grouper nervous necrosis virus (OGNNV) were produced in prokaryotes and their ability to enter Asian sea bass cells was the same as native virus, suggesting that they possess a similar structure to OGNNV. VLPs immunogenicity was then determined by intramuscularly vaccinating Epinephelus coioides at different concentrations (1.5 or 15 μg g^?1 fish body weight, FBW) and immunizing frequencies (administration once, twice and thrice). A single vaccination with the dosage of 1.5 μg g^?1 FBW is enough to provoke high titer antibodies (average 3 fold higher than that of negative control) with strong neutralizing antibody titer as early as 1 week post immunization. Furthermore, quantitative PCR analysis revealed that eleven genes associated with humoral, cellular and innate immunities were up-regulated in the liver, spleen and head kidney at 12 h post immunization, correlating with the early antibody response. In conclusion, we demonstrated that VLP vaccination induced humoral immune responses and activated genes associated with cellular and innate immunity against betanodavirus infection in orange-spotted grouper.     

Pharmacodynamics of amoxicillin against Mannheimia haemolytica and Pasteurella multocida and pharmacokinetic/pharmacodynamic (PK/PD) correlation in sheep

Silicone-made tissue cages were implanted in sheep. Blood serum (SBS) and tissue cage fluid (TCF) samples were collected after amoxicillin intravenous and intramuscular administrations, at the dose of 15 mg/kg. Amoxicillin pharmacodynamics were studied in an artificial culture medium, SBS and TCF with use of a Mannheimia haemolytica and a Pasteurella multocida strain. A concentration-independent antimicrobial activity of amoxicillin was confirmed for levels higher than 0.79–1.75 × MIC. This result favored the use of the percentage of the 24 h dosing interval during which drug levels remain above MIC as the appropriate pharmacokinetic/pharmacodynamic index. The subsequent correlation revealed that intravenous administration could be considered effective against “deep” infections caused by bacteria with MICs < 1 μg/mL or “shallow” infections caused by bacteria with MICs < 0.1 μg/mL. Intramuscular administration could be safely considered effective against both “deep” and “shallow” infections when the MICs of the targeted pathogens are lower than 1 μg/mL.     

Stocking Rate and Fuels Reduction Effects on Beef Cattle Diet Composition and Quality

An experiment was conducted to evaluate the influence of forest fuels reduction on diet quality, botanical composition, relative preference, and foraging efficiency of beef cattle grazing at different stocking rates. A split plot factorial design was used, with whole plots (3 ha) being fuel reduced or no treatment (control), and split plots (1 ha) within whole plots were grazed to three levels of forage utilization; (low) 3 heifers · ha^?1, (moderate) 6 heifers · ha^?1, (high) 9 heifers · ha^?1, with a 48-h grazing duration. Grazing treatments were applied in August of 2005 and 2006. Cattle diet composition and masticate samples were collected during 20-min grazing bouts using six ruminally cannulated cows in each experimental unit. Relative preference indices indicated a strong preference for grass regardless of treatment and stocking rate. Grass consumption was lower in control pastures (P < 0.05) and tended (P < 0.095) to decrease with increased stocking rates. Shrub use was higher in control pastures displaying a quadratic effect (P < 0.05) due to stocking, whereas shrub use increased with stocking rate across all treatments. Cattle grazing control pastures consumed diets higher in crude protein compared to cattle grazing treated pastures (P < 0.05). In vitro dry matter digestibility values were lower (P < 0.05) in control sites and tended (P = 0.10) to decrease with increased stocking rates. In both control and treated pastures, bites per minute and grams consumed per minute declined (P = 0.003) with increased stocking, indicating foraging efficiency of cattle decreases with increased stocking rates. Our data indicated cattle grazing late season grand fir habitat types have a strong preference for grasses regardless of treatment or stocking rate. However, as stocking rate increased in both control and treated pastures, grass consumption decreased, shrub consumption increased, and foraging efficiency decreased.     

Diet selection by calves facing pairs of nutritionally complementary foods

Experimental evidence suggests that ruminant animals are capable of selecting a nutritionally balanced diet (i.e. a diet appropriate to their metabolic needs) from imbalanced but complementary foods. We tested this hypothesis by offering calves different combinations of the same complementary foods. A 5 × 5 Latin square design was used to assess the effect on diet selection of different combinations (termed as food A and food B) of two complementary foods (alfalfa-grass hay and maize grain) offered in separate feed bunks. In Treatment 1, food A and food B had the same composition, 50.0% hay and 50.0% grain (T50:50). In the rest of the treatments, hay–grain combinations in foods A and B were, respectively: 62.5–37.5% and 37.5–62.5% (T63:37); 75.0–25.0% and 25.0–75.0% (T75:25); 87.5–12.5% and 12.5–87.5% (T88:12); and 100.0–0.0% and 0.0–100.0% (T100:0). Daily intake data were analysed through repeated measures analysis. Crude protein intake was similar across treatments (P = 0.28). An increasing tendency (P = 0.01) in food B intake (the food with more grain content) from T50:50 to T100:0, however, led into a decreasing tendency (P < 0.001) in the protein: energy ratio of the diet across treatments. Grain consumption in treatments T75:25, T88:12 and T100:0 was higher than expected, leading to protein levels in the diet below those required for a balanced diet (predicted vs. observed: 19.38 vs. 16.67, P = 0.004; 20.53 vs. 16.10, P = 0.001; and 20.23 vs. 16.79 g kgMW^? 1 d^? 1, P = 0.015, for T75:25, T88:12 and T100:0, respectively). Although calves were potentially able to select a balanced diet in all feeding treatments, they failed when offered high grain foods at choice (75% of maize grain or more). Evolved mechanisms to regulate macronutrient intake under natural conditions may fail to adequately operate in artificial environments involving high-energy density foods (e.g. maize grain).     

Characterization of a monoclonal antibody to Spiroplasma eriocheiris and identification of a motif expressed by the pathogen

Tremor disease (TD) has been found to be a lethal epidemic in Chinese mitten crab during recent years. A new species of spiroplasma, Spiroplasma eriocheiris (S. eriocheiris), was identified as the pathogen causing TD. In order to acquire appropriate biological tools for characterizing this newly discovered pathogen, we developed a new S. eriocheiris specific monoclonal antibody (mAb) 6H7. The antibody showed high binding affinity to S. eriocheiris (K_a = 6.25 × 10⁸ M^?1) and it bound to the adhesin-like protein (ALP41) of S. eriocheiris in Western blot analysis. Screening of a commercially available 12-mer linear peptide library by using 6H7 as bait led to isolation of a consensus sequence (FQGINHYNQMER). The clone displaying this sequence exhibited a significant, dose-dependent binding to 6H7 and inhibited the binding of S. eriocheiris to 6H7, suggesting a similar epitope between the sequence and S. eriocheiris. Homology searches and multiple sequence alignments indicated moderate homology between the consensus peptides and the ALP of S. eriocheiris. Thus, an internal image of 6H7 binding epitope might be buried in ALP41 of S. eriocheiris. In conclusion, 6H7 is a promising mAb to identify S. eriocheiris and the consensus sequence can be used in future research on the characterization of S. eriocheiris and its pathogenesis.     

New multiplex PCR method for the simultaneous diagnosis of the three known species of equine tapeworm

Although several techniques exist for the detection of equine tapeworms in serum and feces, the differential diagnosis of tapeworm infection is usually based on postmortem findings and the morphological identification of eggs in feces. In this study, a multiplex polymerase chain reaction (PCR)-based method for the simultaneuos detection of Anoplocephala magna, Anoplocephala perfoliata and Anoplocephaloides mamillana has been developed and validated. The method simultaneously amplifies hypervariable SSUrRNA gene regions in the three tapeworm species in a single reaction using three pairs of primers, which exclusively amplify the internal transcribed spacer 2 (ITS-2) in each target gene. The method was tested on three types of sample: (a) 1/10, 1/100, 1/500, 1/1000, 1/2000 and 1/5000 dilutions of 70 ng of genomic DNA of the three tapeworm species, (b) DNA extracted from negative aliquots of sediments of negative control fecal samples spiked with 500, 200, 100, 50 and 10 eggs (only for A. magna and A. perfoliata; no A. mamillana eggs available) and (c) DNA extracted from 80, 50, 40, 30, 10 and 1 egg per 2 μl of PCR reaction mix (only for A. magna and A. perfoliata; no A. mamillana eggs available). No amplification was observed against the DNA of Gasterophilus intestinalis, Parascaris equorum and Strongylus vulgaris. The multiplex PCR method emerged as specific for the three tapeworms and was able to identify as few as 50 eggs per fecal sample and as little as 0.7 ng of control genomic DNA obtained from the three species. The method proposed is able to differentiate infections caused by the two most frequent species A. magna or A. perfoliata when the eggs are present in feces and is also able to detect mixed infections by the three cestode species.     

Pharmacokinetics and ex vivo pharmacodynamics of cefquinome in porcine serum and tissue cage fluids

A tissue cage (TC) model was used to evaluate the pharmacokinetics and ex vivo pharmacodynamics of cefquinome after intravenous (IV) and intramuscular (IM) administration to piglets at 2 mg/kg bodyweight. The mean values of area under the concentration–time curve (AUC) were 21.28 (IV) and 21.37 (IM) μg h/mL for serum, and 17.40 (IV) and 16.57 (IM) μg h/mL for TC fluid (TCF), respectively. Values of maximum concentration (C_max) were 6.15 μg/mL (serum) and 1.15 μg/mL (TCF) after IM administration. The elimination half-lives (t_1/2β) in TCF (10.63 h IV and 11.81 h IM) were significantly higher than those in serum (2.33 h IV and 2.30 h IM) (P < 0.05). The values of AUC_TCF/AUC_serum (%) after IV and IM administration were 82.4% and 80.7%, respectively.The ex vivo time-kill curves were established for serum and TCF samples using Escherichia coli ATCC 25922. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration values of cefquinome against E. coli were 0.030 and 0.060 μg/mL in Mueller–Hinton broth, and 0.032 and 0.064 μg/mL in both serum and TCF, respectively. The ex vivo growth inhibition data of TCF after IM administration were fitted to the sigmoid E_max model; AUC_24h/MIC was 35.01 h for bactericidal activity and 44.28 h for virtual eradication, respectively. The findings from this study suggest that cefquinome may be therapeutically effective in diseases of pigs caused by E. coli when used at a dose rate of 1.33 mg/kg administered every 24 h for organisms with MIC₉₀ ⩽ 0.50 μg/mL.