Comparative evaluation of therapeutic efficacy of sulfadiazine-trimethoprim,oxytetracycline, enrofloxacin and florfenicol on Staphylococcus aureus–induced arthritis in broilers

1. Staphylococcus aureus is an important human and veterinary pathogen that causes economic loss in the poultry industry. This study aimed to compare therapeutic efficacy of 4 commonly used antibiotics in poultry on S. aureus–induced arthritis in broilers.

2. Sixty broilers, 8 weeks of age, were assigned at random into 7 groups as follows: (1) negative control (n = 5); (2) vehicle control (n = 5); (3) sulfadiazine-trimethoprim, 250 ml/1000 l drinking water (n = 10); (4) oxytetracycline 20%, 1 mg/l drinking water (n = 10); (5) florfenicol 10%, 1/1000 v/v in drinking water (n = 10); (6) enrofloxacin 10%, 1/1000 v/v in drinking water (n = 10) and (7) positive control (n = 10).

3. Birds in group 2 were injected with 1 ml of sterile TSB medium into the right tibiotarsal joint on d 0 while other birds (except group 1) were challenged with 1 ml of 1.2 × 10¹⁰ CFU/ml suspension of S. aureus bacteria.

4. Antibiotic therapy was started from d 4 post challenge and continued for 5 d. At the end, birds were weighed and clinical severity of arthritis was determined. After blood collection, birds were slaughtered and tibiotarsal and hip joints were evaluated grossly. The content of inflammatory exudates of tibiotarsal joint and the degree of femoral head necrosis were recorded. Mucin clot test and histopathological evaluation were performed on right tibiotarsal joint. Serum interleukin 6 was also assayed.

5. Sulfadiazine-trimethoprim had higher therapeutic efficiency with regard to most of the assayed criteria, whereas none of the antibiotics significantly affected femoral head necrosis and body weight. These data will help clinicians to have better antibiotic choice in field conditions.

     

The effect of different dietary levels of sodium and chloride on performance and blood parameters in goslings (1–28 days of age)

A total of 702 1-day-old male Jiangnan White goslings were used to study the effects of varying levels of Na⁺ and Cl⁻ on their growth performance and blood parameters. In this experiment, goslings were randomly allotted to nine treatments, with six pens per treatment in a 3 × 3 factorial arrangement, and fed diets with three concentrations of added Na⁺ (0.10%, 0.15% and 0.20%) and three concentrations of added Cl⁻ (0.15%, 0.20% and 0.25%). The purpose of this experiment was to investigate the sensitivity of goslings to Na⁺ and Cl⁻ during brooding to determine the appropriate levels of Na⁺ and Cl⁻ in their feed. The results are as follows: (a) Different levels of Na⁺ and Cl⁻ in the diet, especially low Na⁺ and Cl⁻, significantly affected the body weight (BW), average daily gain (ADG) and feed/gain ratio (F/G) of the 28-day-old goslings (p < .05). But Na⁺ × Cl⁻ has no significant effect on water consumption (p > .05). (b) Serum concentrations of urea increased linearly with Na⁺ content (p < .05). Serum concentrations of creatinine (CR) and uric acid (UA) increased linearly with Cl⁻ content (p < .05). (c) Na⁺ × Cl⁻ has a significant impact on the serum sodium (Na) and chlorine (Cl; p < .05). The content of serum Na and Cl increased linearly with increasing levels of Na⁺ and Cl⁻. These results show that low levels of Na⁺ and Cl⁻ had significant adverse effects on the growth of 1- to 28-day-old goslings. The results of the experiment support a recommendation to supplement the diet of goslings with 0.20% Cl⁻ and not <0.15% Na⁺.     

The influence of age and body weight gain on enrofloxacin pharmacokinetics in turkeys—Allometric approach to dose optimization

Enrofloxacin is a concentration-dependent antimicrobial used in bacterial infections in poultry. During a few months of a turkey's life, pharmacokinetics of drugs undergoes substantial changes which may compromise their efficacy due to variability in internal exposure (measured by area under the concentration–time curve, AUC). The aim of this study was to describe the effects of age on the pharmacokinetics of a single intravenous (i.v.) and oral administration of enrofloxacin at a dose of 10 mg/kg to turkeys. It was found that during a 2.5-month-long period of growth from 1.4 to 14.6 kg, the AUC after i.v. administration increased almost threefold due to a significant decrease in the body clearance (from a mean of 0.76–0.28 L hr⁻¹ kg⁻¹). Over the same period, the mean elimination half-life was prolonged from 2.65 to 7.03 hr. Oral administration resulted in a similar trend in pharmacokinetic parameters. For both routes, formation of the major metabolite, ciprofloxacin, was marginal. Protein binding was not age-dependent and never exceeded 50%. Body clearance, volume of distribution and elimination half-life were subjected to an allometric analysis and a novel, nonlinear dosage protocol has been proposed to improve the internal exposure to the drug in different age groups of turkeys.     

The Kümm isolate of Ehrlichia ruminantium: in vitro isolation,propagation and characterization

An effective culture system for Ehrlichia (Cowdria) ruminantium comb. nov. was first established in 1985 and many stocks were subsequently isolated and propagated in vitro. A notable exception, however, was the Kümm isolate that resisted all attempts at in vitro culture until the successful experiment described here. In one experiment white blood cells were harvested from heparinized blood derived from a sheep infected with the Kümm isolate. The cells were added to DH 82 cells and incubated at 37 degrees C. The high metabolic activity of the DH 82 cells necessitated that cell growth be retarded by the addition of cycloheximide. Colonies were first detected 19 days after culture initiation and, once the cultures were established, they could be passaged every 3 days. Bovine and sheep endothelial cells were readily infected with culture supernatant obtained from the infected DH 82 cells. In a further experiment another sheep was infected, using a higher dose of the same batch of Kümm stabilate, and we attempted to infect several different cell lines: these were DH 82 cells, bovine aorta (BA 886) cells, sheep brain endothelial (SBE 189) cells and sheep fibroblastoid cells (E2). Ten days after culture initiation only the E2 cells had become positive for E. ruminantium. Culture supernatant from the first cultured isolate (Kümm-1) was less virulent for mice than that of the second cultured isolate (Kümm-2) which killed all mice. Upon molecular characterization with E. ruminantium 16S probes we found that Kümm-1 hybridized with a Senegal 16S genotype probe, whereas Kümm-2 hybridized only with an Omatjenne 16S genotype probe. The original stabilate used to infect the sheep hybridized with both probes. These results clearly indicate that two different stocks had been isolated in culture.     

The influence of trilostane on steroid hormone metabolism in canine adrenal glands and corpora lutea—an in vitro study

Trilostane is widely used to treat hyperadrenocorticism in dogs. Trilostane competitively inhibits the enzyme 3-beta hydroxysteroid dehydrogenase (3β-HSD), which converts pregnenolone (P5) to progesterone (P4) and dehydroepiandrosterone (DHEA) to androstendione (A4). Although trilostane is frequently used in dogs, the molecular mechanism underlying its effect on canine steroid hormone biosynthesis is still an enigma. Multiple enzymes of 3β–HSD have been found in humans, rats and mice and their presence might explain the contradictory results of studies on the effectiveness of trilostane. We therefore investigated the influence of trilostane on steroid hormone metabolism in dogs by means of an in vitro model. Canine adrenal glands from freshly euthanized dogs and corpora lutea (CL) were incubated with increasing doses of trilostane. Tritiated P5 or DHEA were used as substrates. The resulting radioactive metabolites were extracted, separated by thin layer chromatography and visualized by autoradiography. A wide variety of radioactive metabolites were formed in the adrenal glands and in the CL, indicating high metabolic activity in both tissues. In the adrenal cortex, trilostane influences the P5 metabolism in a dose- and time-dependent manner, while DHEA metabolism and metabolism of both hormones in the CL were unaffected. The results indicate for the first time that there might be more than one enzyme of 3β-HSD present in dogs and that trilostane selectively inhibits P5 conversion to P4 only in the adrenal gland.     

The effect of α-tocopherol,sweet chestnut wood extract and their combination on oxidative stress in vivo and the oxidative stability of meat in broilers

1. This study examined the effect of α-tocopherol (α-T), sweet chestnut wood extract (SCW) and their combination on oxidative stress in vivo and oxidative stability of meat in broilers given diets rich in PUFA.

2. A total of 60 male broilers were individually caged and divided into 6 groups of 10. The C-PALM group received a diet with 7·5% palm fat and the other 5 groups with 7·5% linseed oil. The linseed oil groups were either un-supplemented (C-LIN) or supplemented with α-T or/and SCW as follows: αT-85 (C-LIN diet?+?68?IU vit E as all-rac-α-T/kg), αT-200 (C-LIN diet?+?183?IU vit E as all-rac-α-T/kg), SCW (C-LIN diet?+?3?g SCW/kg) and αT-SCW (C-LIN diet?+?68?IU vit E as all-rac-α-T/kg?+?3?g SCW/kg). Different parameters of oxidative stress were measured.

3. Linseed oil induced DNA fragmentation and malondialdehyde (MDA) formation, while α-T reduced both parameters, and SCW reduced the DNA damage. A combination (αT-SCW) also reduced plasma MDA. Larger antioxidant capacity of lipid soluble compounds were recorded in groups αT-85, αT-200 and αT-SCW than in the controls but there were no differences between these groups in antioxidant enzymes and total antioxidant status. A combination (αT-SCW) increased tocopherol concentrations in breast muscle and in comparison to the C-LIN group MDA concentrations were reduced in groups αT-85, αT-200 and αT-SCW.

4. It can be concluded that neither of the α-T concentrations were able to prevent all the negative effects of lipid oxidation in vivo and only high concentrations of α-T improved the stability of meat. With the exception of DNA damage, SCW had no impact on in vivo and in vitro measured markers of oxidative stress but may have a sparing or regenerating effect on α-T.     

Effects of lycopene on in vitro quality and lipid peroxidation in refrigerated and cryopreserved turkey spermatozoa

1. The effects of lycopene-enriched extenders on the in vitro quality of turkey semen including lipid peroxidation were examined after chilled and frozen storage.

2. Five pools of semen diluted in extenders containing 0, 0·05 or 0·1?mg/ml of lycopene were stored at 5°C for 48?h or cryopreserved as pellets and the following variables determined in fresh samples and samples stored chilled or frozen: sperm motility, viability, osmotic resistance, DNA integrity and lipid peroxidation (as malonaldehyde production).

3. Semen quality was generally compromised after storage, especially post-freezing. However, in the presence of the highest dose of lycopene, both the viability and osmotic-resistance of chilled spermatozoa and the DNA integrity of frozen spermatozoa were similar to those of fresh spermatozoa.

4. Greater lipid peroxidation was detected in refrigerated compared to fresh or cryopreserved spermatozoa. However, spermatozoa chilled in lycopene-enriched extenders showed significantly lower malonaldehyde levels than those chilled without lycopene, while the addition of lycopene to the freezing medium served to maintain the lipid peroxidation levels observed in fresh semen.

5. In conclusion, the presence of lycopene in the extender improved the survival of turkey spermatozoa after liquid-storage and protected DNA integrity against cryodamage. The beneficial effects of lycopene observed could be related to its capacity to diminish sperm lipid peroxidation during refrigeration or cryopreservation.     

Growth performance of Hubbard × Cobb 500 and Ross × Ross 708 male broilers fed diets varying in apparent metabolizable energy from 14 to 28 days of age

The cost of energy-contributing ingredients has increased over the last several years. Defining minimum dietary energy specifications for broilers is a strategy to mitigate high diet cost. Two experiments were conducted to evaluate the effects of diets with graded concentrations of AME_n fed to Hubbard × Cobb 500 (experiment 1) and Ross × Ross 708 (experiment 2) male broiler chicks from 14 to 28 d of age. Both experiments were identical in experimental procedures with the exception of genetic strain. Broilers were fed a common corn-soybean meal-based starter diet from 1 to 13 d of age. At 14 d of age, dietary treatments were provided with 1 of 6 concentrations of AME_n (3,000, 3,030, 3,060, 3,090, 3,120, and 3,150 kcal/kg). In experiment 1, BW gain, feed intake, and FCR of Hubbard × Cobb 500 broilers decreased linearly with increasing AME_n. Linear broken-line response of FCR was estimated at 3,062 kcal of AME_n/kg based on broken-line methodology. In experiment 2, Ross × Ross 708 broilers did not respond to graded concentrations of energy, as indicated by linear and quadratic regression of BW gain, feed intake, caloric conversion, and FCR data. Therefore, Hubbard × Cobb 500 male broilers respond to AME_n from 14 to 28 d of age and may be fed diets formulated at 3,062 kcal of AME_n/kg without loss of performance. Conversely, growth performance of Ross × Ross 708 male broilers was not affected by varying AME_n (3,000 to 3,150 kcal/kg) from 14 to 28 d of age. In addition, the response to AME_n may not be as pronounced with the modern broiler compared with broilers used in previous research.     

Bordetella bronchiseptica experimental model development in pigs and efficacy evaluation of a single intramuscular injection of gamithromycin (Zactran® for Swine) against Bordetella bronchiseptica-associated respiratory disease in experimentally infected piglets

In the Bordetella bronchiseptica infection model development study, twenty-eight piglets were inoculated with B. bronchiseptica strain of either canine (10⁹ CFU/ml) or swine (10⁸ and 10⁹ CFU/ml) origin; swine origin strain at 10⁹ CFU/ml was chosen for the efficacy assessment study due to higher incidence and severity of gross and histopathological lesions compared with other strains. To assess efficacy of gamithromycin against B. bronchiseptica, forty piglets were experimentally inoculated on Day 0 and clinical signs were scored as per severity. Animals were then treated either with gamithromycin or saline on Day 3. The Global Clinical Scores in gamithromycin-treated group were consistently lower than the saline-treated control group from Day 4 onwards and were 0 and 40 in the gamithromycin-treated and saline-treated control groups, respectively, on Day 6. Severity and frequency of gross and histopathological observations were significantly lower in gamithromycin-treated animals compared with saline-treated controls. The efficacy of Zactran® for Swine at the label dose for the treatment of B. bronchiseptica–associated respiratory disease was demonstrated based on the faster reduction in clinical signs as early as 1 day post-gamithromycin treatment and based on the significant difference in the severity of macroscopic and microscopic lung lesions 10 days post-gamithromycin treatment.     

Antimicrobial susceptibility and molecular subtypes of Staphylococcus aureus isolated from pig tonsils and cow’s milk in China

This study investigated and compared the antimicrobial resistance patterns and ribotypes of Staphylococcus aureus isolated from pig tonsils and cow’s milk in China. A total of 90 isolates of S. aureus was included: 42 strains were isolated from tonsils of pigs and 48 from half-udder milk. The broth microdilution method and the double-disc diffusion test (D test) were used for antimicrobial susceptibility testing. The mecA gene for methicillin-resistant S. aureus (MRSA) and the ermA, ermB, ermC, and msrA genes for erythromycin-resistant strains were detected by polymerase chain reaction (PCR). The isolates were ribotyped with the Riboprinter system. The highest frequency of resistance was observed with clindamycin (91.1%), followed by penicillin (90.0%), and erythromycin (85.6%). All strains were susceptible to vancomycin and trimethoprim-sulfamethoxazole. The D test showed that 54.5% (42/77) of erythromycin-resistant isolates had the constitutive resistance phenotype and 45.5% (35/77) had the inducible resistance phenotype to clindamycin. A higher proportion of resistance to cephalosporins, macrolides, fluoroquinolones, and pleuromutilins was observed in pig isolates than in milk isolates (P < 0.05). The mecA gene was detected in all MRSA isolates; 89.6% of erythromycin-resistant strains harbored the ermC gene and 16.9% harbored the ermB gene. A total of 35 different ribogroups was found among the isolates investigated; 83.3% of pig strains belonged to 1 cluster with a similarity coefficient of 0.84. In contrast, 3 main clusters were observed among 68.8% of milk strains, which indicates a high degree of host specificity.     

The distribution and spreading pattern of Dermacentor reticulatus over its threshold area in the Czech Republic--how much is range of this vector expanding?

Host-seeking Dermacentor reticulatus ticks were detected by flagging method at 46 localities at south-east part of the Czech Republic, in the basins of rivers Morava and Dyje. Exact north-west distribution limits of D. reticulatus were defined in this area for the first time. Detailed prediction map of probabilities of D. reticulatus occurrence was obtained using GIS analysis. Spatial model delimited a south-north gradient in probability across the studied area, with highest probabilities above 0.8 in its southernmost part. Abundance of D. reticulatus varied markedly between localities in interval 0.33-222 of ticks per flag per hour. The highest abundances were in flooded areas at lower streams, towards upper streams abundance and density of these ticks decreased. Females prevailed in samples with population sex ratio of 0.413, significantly deviating from parity. Larvae and nymphs of this species were not detected by flagging. Although D. reticulatus range expansion probably did not reach such a degree as reported in other countries, these ticks became very abundant in some parts of studied area. Since spreading of vector-borne diseases became a problem in Europe, the knowledge of their exact recent geographic ranges is important for future modelling of their shift predictability.     

Evaluation of clinical safety and anthelmintic efficacy of aurixazole administed orally at 24 mg/kg in cattle

The current study evaluated, in vivo, the clinical safety and the anthelmintic efficacy of 24% aurixazole (24 mg/kg), administered orally, in bovines. Two experiments were conducted: the first one evaluating the clinical safety of 24% aurixazole (24 mg/kg) in cattle, and a second one evaluating the anthelmintic efficacy of aurixazole (24 mg/kg) against gastrointestinal nematodes on naturally infected cattle. Based on the results of clinical safety, no alterations on clinical and haematological signs and on the biochemical values obtained in animals treated orally with aurixazole 24 mg/kg were observed. Regarding the results of reduction or efficacy, obtained by eggs per gram of faeces (EPG) counts, the formulation of aurixazole reached values superior to 99% (arithmetic means) in all post-treatment dates. In two occasions, this formulation reached maximum efficacy (100%). Comparing these results with the reduction percentages obtained by EPG counts, it is possible to verify that the values obtained by all three formulations were compatible with the efficacy results. Aurixazole reached maximum efficacy (100%) against Haemonchus placei, Cooperia spatulata and Oesophagostomum radiatum. Against Cooperia punctata, this formulation reached an efficacy index of 99.99%. Regarding aurixazole, no specific trials were conducted on the field in order to evaluate the behaviour of this molecule against helminths that are resistant to other molecules, specially isolated levamisole and disophenolat. Due to this fact, future studies will be necessary to assess the effectiveness of aurixazole against strains of nematodes that are resistant to levamisole and disophenolat, but the results of clinical safety and efficacy described in this study allow us to conclude that the aurixazole molecule, concomitantly with other measures and orally administered formulations, can be another important tool in the control of nematodes parasitizing bovines.     

Involvement of tannins and flavonoids in the in vitro effects of Newbouldia laevis and Zanthoxylum zanthoxyloïdes extracts on the exsheathment of third-stage infective larvae of gastrointestinal nematodes

The present study aimed at examining the possible role of tannins and flavonoids on the in vitro anthelmintic properties of the extracts of two plants from the southern area of Western Africa, i.e. Newbouldia laevis and Zanthoxylum zanthoxylo?des. Extracts of the two plants were prepared by use of acetone/water (70/30) and their anthelmintic activity was measured by use of the larval exsheathment inhibition assay (LEIA) applied on the abomasal species, Haemonchus contortus and the intestinal species Trichostrongylus colubriformis. Three concentrations of extracts were evaluated to examine the possible dose effect. In addition, the possible involvement of tannins and flavonoids was examined by comparing the levels of inhibition of larval exsheathment obtained with the same extracts, after of not addition of PVPP which forms complexes with these compounds. The results indicate significant effects with both plants and both nematode species. In the range of concentrations examined, the results were dose-dependent for N. laevis extracts but not for Z. zanthoxylo?des because the three doses applied provoked a similar highly significant inhibition whatever the tested dose. The use of PVPP indicated for both plant and nematode species, that tannins and flavonoids are involved partly in the effect but that some other biochemical compounds were also involved in both plants.     

The measurement of three cytokine transcripts in naïve and sensitized ovine peripheral blood mononuclear cells following in vitro stimulation with bluetongue virus serotype-23

Bluetongue (BT), a serious disease of sheep and some wild ruminants, is caused by bluetongue virus (BTV), a member of the family, Reoviridae. The current research thrust for controlling BT is on development of efficient vaccines, necessitating clear understanding of ovine immunology. At present, comparative studies on cytokine gene expression profiles of na?ve and BTV-sensitized peripheral blood mononuclear cells (PBMC) in sheep have not been clearly understood. In the present study, PBMC from na?ve and BEI-inactivated-saponin-adjuvanted BTV-1 vaccinated sheep were stimulated in vitro with heterologous BTV-23. At various intervals, RT-qPCR was carried out to estimate cytokine (interferon-gamma, interleukin-12 and interleukin-2) mRNA expressions that are linked to cell-mediated immunity. The results showed that PBMC cytokine profiles were relatively increased both temporally and quantitatively in immunized sheep PBMC compared to na?ve ones, suggesting that BTV-1 vaccination may prime immune system that can cross-react with BTV-23 antigens.     

Comparative efficacy of a spot-on formulation containing emodepside and praziquantel (Profender®, Bayer) and praziquantel and pyrantel oral tablets (Drontal® for Cats) against experimental Ancylostoma ceylanicum infections in cats

Ancylostoma ceylanicum is a common zoonotic hookworm of dogs and cats throughout Asia and has also been reported to occur within the Australasian region. The aim of this study to was to determine the efficacy of a spot-on formulation containing emodepside and praziquantel (Profender^®, Bayer) and praziquantel and pyrantel oral tablets (Drontal^® for Cats, Bayer) against experimental A. ceylanicum infections in cats.Twenty-four kittens were each subcutaneously injected with 100 infective third-stage larvae of A. ceylanicum. Kittens were stratified by egg count and randomly allocated equally into control and two treatment groups. The first group were treated with emodepside 2.1%/praziquantel 8.6% (Profender®, Bayer) at the recommended label dose. The second group was treated with 80 mg pyrantel and 20 mg praziquantel (Drontal^® for Cats, Bayer) at the recommended label dose. The kittens in the control group were not treated. Egg counts were performed daily until the end of the study period and compared for the treated and control groups. No eggs were detected in the treated group of kittens within 4 days of treatment and faecal samples from this group remained negative throughout the rest of the study, resulting in a treatment efficacy (egg reduction) of 100% (P < 0.0001). The egg counts remained high (993 ± 666 epg) in the untreated control group for the rest of the study period. This study demonstrated that both combination products containing topical emodepside/praziquantel (Profender^®, Bayer) and praziquantel/pyrantel oral tablets (Drontal^® for Cats, Bayer) given at the recommended dose is highly effective against infection with A. ceylanicum in cats.