Comparison of Different Agar Diffusion Methods for the Detection of Antimicrobial Residues in Slaughter Animals

The different agar diffusion methods were compared using antibiotic and sulphonamide-impregnated filter-paper discs and the kidneys of healthy and emergency-slaughtered pigs and cows after slaughter.No method used seemed to be sensitive to all antimicrobial drugs preimpregnated onto discs. Tetracycline yielded a greater zone of inhibition at pH 6 than at pH 8 and aminoglycosides, erythromycin, polymyxin B and lin cornycin at pH 8 than at pH 6. It therefore seems necessary to use different pHs (6 and 8). The addition of trimethoprim to the medium is necessary for the detection of sulphonamides. Bacillus subtilis BGA used as the test organism was more sensitive to sulphonamides on the “Test agar for the inhibitor test” containing trimethoprim than on the “Iso-Sensitest agar” also containing trimethoprim. The addition of trimethoprim to “Test agar for the inhibitor test” is recommended at pH 8 but not at pH 6 because false-positive cases (with inhibition zones > 2 mm) were observed at pH 6 with trimethoprim on the kidneys of healthy pigs.     

Distribution of porcine monocytes in different lymphoid tissues and the lungs during experimental Actinobacillus pleuropneumoniae infection and the role of chemokines

Monocytes play an essential role in the defense against bacterial pathogens. Bone marrow (BM) and peripheral blood (PB) monocytes in pigs consist of the main “steady-state” subpopulations: CD14^hi/CD163^-/SLA-DR^- and CD14^low/CD163⁺/SLA-DR⁺. During inflammation, the subpopulation of “inflammatory” monocytes expressing very high levels of CD163, but lacking the SLA-DR molecule (being CD14^low/CD163⁺/SLA-DR^-) appears in the BM and PB and replaces the CD14^low/CD163⁺/SLA-DR⁺ subpopulation. However, current knowledge of monocyte migration into inflamed tissues in pigs is limited. The aim of the present study was to evaluate the distribution of “inflammatory” CD14^low/CD163⁺/SLA-DR^- monocytes during experimental inflammation induced by Actinobacillus pleuropneumoniae (APP) and a possible role for chemokines in attracting “inflammatory” CD14^low/CD163⁺/SLA-DR^- monocytes into the tissues. Monocyte subpopulations were detected by flow cytometry. Chemokines and chemokine receptors were detected by RT-qPCR. The “steady-state” monocytes were found in the BM, PB, spleen and lungs of control pigs. After APP-infection, “inflammatory” monocytes replaced the “steady-state” subpopulation in BM, PB, spleen and moreover, they appeared in an unaffected area, demarcation zone and necrotic area of the lungs and in tracheobronchial lymph nodes. They did not appear in mesenteric lymph nodes. Levels of mRNA for various chemokines with their appropriate receptors were found to be elevated in BM (CCL3-CCR1/CCR5, CCL8-CCR2/CCR5, CCL19-CCR7), necrotic area of the lungs (CCL3-CCR1, CCL5-CCR1/CCR3, CCL11-CCR3, CCL22/CCR4) and tracheobronchial lymph nodes (CCL3-CCR1) and therefore they could play a role in attracting monocytes into inflamed tissues. In conclusion, “inflammatory” monocytes appear in different lymphoid tissues and the lungs after APP infection in pigs. Various chemokines could drive this process.     

Clinico-Pathologic Findings in Young Pigs Fed Different Levels of Selenium,Vitamin E and Antioxidant

A randomized, blocked 2³ factorial experiment was conducted with 48 young pigs. The treatment factors were: 2 levels of selenium (55 and 115 µg/kg), 2 levels of vitamin E (3 and 53 mg/kg) and 2 levels of the antioxidant feed additive Ethoxyquin (0 and 150 mg/kg). All pigs were kept in single pens and fed ad libitum throughout the experimental period of 9 weeks, i.e. from 3 to 12 weeks of age.Plasma, heart, liver and muscle Se levels as well as whole blood glutathione peroxidase activity (EC 1.11.1.9 GSH-Px) were significantly higher in pigs given a dietary supplement of Se than in pigs given no supplement of Se (P ≤ 0.001). The Se-supplemented pigs showed a tendency to lower mean serum transaminase activity (ASAT and ALAT) than unsupplemented pigs, but the influence was significant (P ≤ 0.05) only for the ALAT activity.Blood vit. E levels were higher for pigs receiving a supplement of vit. E than for unsupplemented pigs (P ≤ 0.001), and so was the resistance of red blood cells against lipid peroxidation (ELP), as expressed by lower ELP values.There were no effects of Ethoxyquin supplementation on the biochemical variables included in the study.The histological examination of heart muscle showed that the score for changes was negatively influenced by both Se and vit. E supplement (P ≤ 0.001) and to some extent also by Ethoxyquin supplement (P ≤ 0.05). The histological picture of m. long dorsi was influenced only by the vit. E supplement (P ≤ 0.01). No histological changes were found in the liver in this study. There were inverse relationships between whole blood GSH-Px defluorescence time and blood Se, and between ELP and whole blood vit. E (P ≤ 0.001).     

Glutathione Peroxidase Activity and Erythrocyte Lipid Peroxidation as Indices of Selenium and Vitamin E Status in Young Pigs

A randomized, blocked 2³ factorial experiment was conducted with 48 pigs from sows fed a diet low in selenium and vitamin E. From 3 to 12 weeks of age the piglets were kept in single pens and fed a basic diet consisting mostly of barley, dried skim milk, soybean meal and dried yeast, and containing 55 µg selenium and 3 mg vitamin E per kg. The treatment factors — i.e. feed supplements — were 2 levels of Se (nil, 60 µg/kg), 2 levels of vitamin E (nil, 50 mg/kg), and 2 levels of the feed antioxidant ethoxyquin (nil, 150 mg/kg). Blood samples, collected at termination of the experiment, were examined for glutathione peroxidase activity (GSH-Px) and resistance against erythrocyte lipid peroxidation (ELP) to evaluate Se and vitamin E status, respectively. Analysis of variance showed the GSH-Px activity to be litter-dependent (P < 0.001) and influenced by selenium supplementation (P < 0.001) but not by the other supplements or by interactions between supplements. Resistance against ELP was influenced only by vitamin E supplementation (P < 0.001). GSH-Px and ELP thus seem to be valuable and simple methods for evaluating, respectively, Se status and vitamin E status in growing pigs.     

Hyperostosis in Newborn Pigs

Hyperostosis “thick legs” is reported in a case involving 7 of a litter of 11 purebred Landrace piglets. The 12 limbs involved showed variable degrees of thickening and immobility; tension and fixation of the skin. Necropsy revealed extensive fibrous connective tissue, enlargement and edema of associated lymph nodes and hypoplasia of muscle groups. Radiographs revealed osteodystrophia along the shafts of long bones. The case is compared to previous reports of infantile cortical hyperostosis in children and “thick legs” in pigs.     

Nitrofuran Production Efficiency In Chickens

Two nitrofuran feed additives, 0.011% nihydrazone and a combination of 0.0055% nitrofurazone and 0.0008% furazolidone, improved weight gains and feed conversions in chickens with “air sac infection.” Both nitrofurans caused a significant reduction in the total chickens condemned at the dressing plant from this disease, but nihydrazone gave the best results.

Nihydrazone^*, a new nitrofuran feed additive for chickens, was found by Wolfgang et al. (1) to be effective against coccidiosis due to Eimeria tenella and E. necatrix. In chickens nihydrazone was shown by Edgar et al. (2) to result in fewer chickens condemned from “air sac infection” than with any other drug used. Rosenberg et al. (3) found nihydrazone caused significant reduction in condemnations due to this disease. Cosgrove (4) showed that nihydrazone prevented an outbreak of cecal coccidiosis, reduced the incidence of “air sac infection,” improved weight gains, feed conversions and livability. Bierer (5) found nihydrazone active against fowl typhoid.

Harwood et al. (6) reported bifuran^** effective against E. tenella and E. necatrix coccidiosis and it has been used commercially for this purpose. Bierer (5) (7) found Bifuran^*** active in prevention of pullorum disease and fowl typhoid in chicks.

The mode of action of nihydrazone and nitrofurazone against E. tenella coccidiosis was shown by Johnson and Van Ryzin (8).

This production efficiency study was undertaken to evaluate nihydrazone and bifuran in the presence of “air sac infection” and concomitant diseases under field conditions. Camden (9) states that the only satisfactory test of a drug is the performance it gives under field conditions.

Nihydrazone (1) (5) and Bifuran (5) (6) have both antibacterial and antiprotozoal activity and under field conditions, normal densities of bacterial and other parasitic organisms are encountered (9). Thus a coccidiostat having antibacterial and antiprotozoal properties is desirable (7).

     

The Epithelium Covering Peyer’s Patches in Young Milk-Fed Calves: An Ultrastructural and Enzyme Histochemical Investigation

Between the ordinary villi over Peyer’s patches there are small domes or “pseudovilli” caused by bulges in the lymphoid tissue. These “pseudovilli” were studied in 5 healthy milk-fed, about 3-week-old, pre-ruminant calves. Scanning electron microscopy revealed that the “pseudovilli” were covered by a specialized follicle associated epithelium (FAE). The FAE had poorly developed microvilli and often extensive folding of the cell surface close to the cell borders. By transmission electron microscopy the tips of the marginal folds of the FAE seemed to fuse, probably in the process of enfolding bulk material from the lumen. The FAE apical cytoplasm contained numerous thick-walled and bristle-coated invaginations, tubules and vesicles indicative of micropinocytosis. Multivesicular bodies and large vacuoles were frequent. Indications of extracellular unloading of residual bodies were found. Intraepithelial lymphocytes tended to group together, and some were rich in rough endoplasmic reticulum. Enzyme histochemistry showed weak reactions of adenosine triphosphate splitting enzyme and aminopeptidase in the FAE luminal cell border. Cytoplasmic acid phosrphatase showed a marked basal-apical decrease along the “pseudovillus” probably caused by the onset of endocytosis. The results of this study appear compatible with the concept that the FAE takes up macromolecules from the lumen by pinocytosis and sensitizes lymphocytes.     

Glasser's Disease of Swine Produced by the Intratracheal Inoculation of Haemophilus suis

The intracheal inoculation of pigs with Haemophilus suis led to the production of Glasser's disease at every attempt without significant pulmonary involvement. Isolation of this organism from the experimental animals was possible only in the acute phase of the disease.

The indirect fluorescent antibody technique when applied to frozen sections of tissues obtained from the experimentally infected pigs at autopsy, revealed a few rod forms but mostly “round bodies” of H. suis in animals from which the organism was isolated, and “round bodies” only in the pigs from which the organism was not isolated.

Attention is drawn to the similarities between the lesions caused by H. suis and Mycoplasma hyorhinis, and to the confusion which may result therefrom. It is stressed that the laboratory diagnosis of these two diseases is complicated by the fact that both agents may not be isolated on the media commonly used in diagnostic laboratories. Both organisms necessitate the use of special media where the clinical and autopsy results indicate polyserositis and arthritis.

     

Flight Distance and Avoidance Score in Thoroughbred Foals and Yearlings and the Relationship with Handling Frequency in the Young Ages

Flight and avoidance reactions from human were examined using 168 postweaning Thoroughbred foals in 22 breeding farms. Further 114 yearlings of 168 foals were tested in the following summer. The foal handlings by the stabler were asked in questionnaire. The relationship between the behavioural reactions and the foal handling frequencies was analyzed. The flight reaction was estimated as the distance from the animal to a stranger when the animal began to flight away from his approach. The avoidance scores were set up from (1) (not resistant) to (5) (touch rejection) from human touching. In the stabler questionnaire, handling frequencies of “body brushing”, “rectal temperature measurement”, “hoof cleaning”, and “stall cleaning” in the early nursing period were asked. The handling frequencies were scored from (1) (not done) to (5) (every day). In the preliminary test, a measurement reliability of the flight distance and the avoidance score was confirmed. The mean flight distances were 0.56 m and 0.27 m in the postweaning foals and the yearlings, respectively. Touch-avoidance scores of the highest frequency were (3) and (2) in the postweaning foals and the yearlings, respectively. As the results of Spearman’s rank-correlation analysis, “body brushing” showed highly negative relationships with “flight distance” (ρ=–0.31, P<0.001) and “avoidance score” (ρ=–0.37, P<0.001) in the postweaning foals. In the yearlings, “hoof cleaning” also showed significantly negative relationships with these behavioural indices (ρ=–0.24, P<0.01; ρ=–0.22, P<0.01, respectively).     

Study of serum drug levels in calves following intramuscular administration of three tetracycline drug preparations

Three antibiotic formulations, oxytetracycline (A) in propylene glycol and oxytetracycline (B) in polyvinyl pyrrolidine and pyrrolidino-methyltetracycline in an oil suspension were given to calves by the intramuscular route. Only oxytetracycline (A) appeared to cause much pain after injection.

The half-time elimination (t_½cl) for oxytetracycline (A) was 14.000 ± 3.580 h for oxytetracycline (B) 10.290 ±5.159 h and for pyrrolidinomethyltetracycline 8.160 ± 0.920 h. The rate of elimination `beta slope” for oxytetracycline (A) was 0.052 ± 0.012 h⁻¹ for oxytetracycline (B) 0.077 ± 0.261 h⁻¹ and for pyrrolidinomethyltetracycline 0.086 ± 0.010 h⁻¹. The Y intercept of the “beta” elimination slope C_os (μg/mL) for oxytetracycline (A) was 2.490 ± 1.040, for oxytetracycline (B) 3.463 ± 1.874 and for pyrrolidinomethyltetracycline 2.852 ± 1.360.

Pyrrolidinomethyltetracycline appeared to have a two component elimination curve, however, only the “beta slope” was used for the calculations.

     

Changes in the predicted function of the rumen bacterial community of Japanese Black beef cattle during the fattening stages according to Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses

We investigated changes in the predicted functions of the rumen bacterial community in Japanese Black beef cattle during fattening. Nine cattle were fed a high-concentrate diet during the early, middle, and late fattening stages consecutively (10–14, 15–22, and 23–30 months of age, respectively). The rumen fluid and solid samples collected at each stage were subjected to sequencing analyses. The sequencing results were clustered and classified into operational taxonomic units (OTUs). Representative sequences and a raw counting table for each OTU were submitted to the Piphillin website. The predicted functions were revealed by the Kyoto Encyclopedia of Genes and Genomes database as the ratio of the total sequence. In the early stage, “Biosynthesis of secondary metabolites” was significantly higher in the fluid fraction than in the solid fraction. “Two-component system” in the middle stage was significantly lower and “Purine metabolism” in the late stage was significantly higher in the fluid fraction than those in the solid fraction. The fluid fraction was significantly correlated with acetic acid, propionic acid, and bacterial metabolism, such as “Biosynthesis of secondary metabolites” and “Sugar metabolism.” Moreover, the solid fraction was correlated with “Purine metabolism” and “Biosynthesis of secondary metabolism”. These results suggest that the rumen bacterial community in Japanese Black beef cattle adapts to changes in rumen conditions by altering their functions in response to a long-term high-grain diet.     

Effects of acepromazine or methadone on midazolam-induced behavioral reactions in dogs

This study evaluated whether acepromazine or methadone reduced behavioral parameters, overall excitement, and activity associated with midazolam administration to healthy dogs. Dogs received midazolam (M) alone [M: 0.25 mg/kg body weight (BW)] or with methadone (MM) (MM: 0.75 mg/kg BW) or acepromazine (MA) (MA: 0.03 mg/kg BW) or saline (S) solution alone, all intramuscularly. Two blinded observers evaluated behavioral parameters using video recordings 30 min before and after injection of drugs. Accelerometery was used to evaluate “total activity counts” (TAC) at baseline and post-treatment. Post-treatment excitement scores were significantly higher in M and MA compared to baseline, M and MM compared to S, and M compared to MA. Behavioral parameters showed significantly higher proportions of “pacing” post-treatment in all groups receiving midazolam, and “restlessness,” “chewing/licking,” and “sniffing” in M. No significant differences were found for TAC at baseline and post-treatment. Midazolam-induced paradoxical behavioral changes (excitation, panting, pacing, restlessness, licking/chewing, and vocalization) were not prevented by acepromazine or methadone in healthy dogs.     

Attempts to Produce Experimental Edema Disease in Swine by Parenterally Injecting Escherichia Coli Serotype 0139:K82:H1

Twenty-two pigs were inoculated parenterally with various E. coli 0139:K82:H1 preparations. Clinical signs of disease in pigs injected with freeze-thaw extract consisted of early listlessness, diarrhea and, later, hyperirritability of varying intensity in some animals.

Hemorrhagic gastroenteritis involving the duodenum, spiral colon and the fundic portion of the stomach, and ulceration of the fundic stomach were observed at post-mortem examination of pigs inoculated parenterallly with living culture or freeze-thaw extract. No significant lesions were observed in pigs inoculated with ultrasonic or hypotonic acid-saline extract.

In pigs injected with living culture or freeze-thaw extract, the histological alterations consisted of moderate perivascular edema of the brain, marked hepatic parenchymal cell degeneration, hepatic subserosal edema and “toxic” lymph nodes, when compared to the control group.

     

Pampas deer (Ozotoceros bezoarticus) courtship and mating behavior

Background

Pampas deer, Ozotoceros bezoarticus (Linnaeus 1758), is a South American grazing deer categorized as "near threatened". However, knowledge about pampas deer behavior including courtship and mating is scarce and incomplete. The aim of this study was to characterize the courtship and mating behavior of the pampas deer (Ozotoceros bezoarticus), an endangered species from South America.

Methods

We performed focal observations of 5 males allocated at the Estación de Cría de Fauna Autóctona Cerro Pan de Azúcar, Uruguay, 4 times a day from 5 to 20 minutes each time on a daily basis from February to May. During that period we recorded all courtship and mating behaviors, as well as quantified the frequency of the specific behaviors shown. As mating were rarely observed, we recorded that behavior when it was observed in the context of other studies performed in the same population during the following 2 years.

Results

During the observation period we recorded 928 courtships and 5 mating periods. In addition, we recorded 10 more matings performed during other studies, totaling 15. The duration of each mating calculated from the 15 recordings was 3.9 ± 0.4 s, and the total period of female receptivity (from first to last mating acceptance) was 8.2 ± 1.1 min. Main observed courtship behaviors in males were “chase” and “ostentation”, while the most observed close to mating were “chinning”, “raised head” and “anogenital sniffing”. The most observed behaviors in females during the mating period were “vulva exhibition” and “move away”.

Conclusion

This is the first detailed report in pampas deer mating behavior. Estrus lasted only 8 min accepting only 3 short copulations per estrus. However, female behavior during courtship can be characterized as highly proceptive.     

The Cause and Prevention of Sacroiliac Arthrosis in the Standardbred Horse: A Theoretical Study

Sacroiliac arthrosis is a major cause of “hiking” lameness (often, so-called “stifle lameness”) in Standardbred horses. The cause of this arthrosis is found in the moment forces generated as horses negotiate unbanked and underbanked turns and in the forces generated on cambered racing surfaces. The prediction is that such lameness could be virtually eliminated if racetrack and training track turns were properly banked and camber held to a minimum.     

Bovine Viral Diarrhea Virus in Swine: Characteristics of Virus Recovered from Naturally and Experimentally Infected Swine

A noncytopathogenic field strain of bovine viral diarrhea virus (BVDV) was isolated from an Iowa farm brood sow and from her hysterectomy-derived, colostrum-deprived (HDCD) piglets. This field isolant was fully virulent for a neonatal calf. The NADL strain of BVDV was passaged through a series of HDCD piglets with no resultant loss of virulence for neonatal calves. Most of the BVD viral isolants recovered from pigs had been changed from a cytopathogenic biotype to a noncytopathogenic biotype. Circumstantial evidence points to swine as “carrier” hosts of BVDV.     

Experimental model of enterotoxigenic Escherichia coli infection in pigs: potential for an early recognition of colibacillosis by monitoring of behavior

The hypothesis that altered behavior is a sign for an early recognition of disease was tested. The experiment was conducted to evaluate the behavioral patterns of pigs in a model of postweaning colibacillosis. Twenty-five weaned pigs (from a herd that was previously found to be highly susceptible to F4⁺ Escherichia coli strains) were randomly assigned into 5 groups, kept in isolated pens under the controlled ambiental conditions. One day after weaning, the pigs from three groups were intragastrically inoculated (via orogastric tube) with either F4ac⁺ (1466 or 2407) or F4⁻ (1467) nonenterotoxigenic E. coli (non-ETEC) strains, respectively. The pigs from the fourth group were inoculated with F4ac⁺ ETEC strain M1823 and the remaining 5 pigs that received broth containing 1.2% sodium bicarbonate were kept as noninoculated controls. The pigs were examined daily and the frequency and duration of their behavioral patterns, such as eating, drinking, lying, standing, urinating, defecating, rooting and playing were monitored for 300 h during a period of 10 days. In this model, three conditions were also observed in F4-susceptible pigs: (1) acute fatal diarrheal disease; (2) moderate diarrhea and weight loss and (3) no diarrhea and weight loss. The incidence (both frequency and duration) of defecating was significantly higher (P<0.05) in pigs inoculated with F4ac⁺ ETEC strain M1823 as compared to that of noninoculated (control) pigs. Pigs inoculated with F4ac⁺ non-ETEC strain 1466 had a significantly lower frequency of eating (P<0.05) and frequency/duration of drinking (P<0.05) than did the controls. The 1466-inoculated pigs, had an increased diarrhea score, but frequency/duration of defecating was not significantly different. Pigs inoculated with F4ac⁺ non-ETEC strain 2407 spent more time in lying (P<0.05) than did noninoculated pigs. Conversely, the pigs that received F4⁻ non-ETEC strain 1467 laid shorter (P<0.05) and ate/drank less frequently (P<0.05) than the controls. It was concluded that the changed occurrence of defecating and eating in pigs that were inoculated with either F4ac⁺ ETEC (M1823) or non-ETEC (1466) strain, respectively, was consistent with the pending clinical disease, i.e. postweaning colibacillosis.     

Target Group Segmentation in the Horse Buyers’ Market against the Background of Equestrian Experience

Whereas in former times horses were reserved primarily for people involved in agriculture, elite equestrians or the military, nowadays equestrian sport has become an activity for people with a wide variety of backgrounds. However, as more and more people become involved with equestrian sport today, the knowledge concerning animal husbandry in general is diminishing due to an alienation from agricultural themes in modern societies. As a consequence, this development affects both riding ability and the appraisal of horses, especially with respect to the purchase of horses. In order to analyse which factors influence purchase decisions in the horse market in conjunction with equestrian experience, 739 horse riders were surveyed on their purchase behaviour in this study. Using cluster analysis, a typology was generated that provides a differentiated picture of the preferences of the various rider groups. Three clusters were distinguished: the “amateurs”, the “experienced” and the “experts”. Taking personal horse riding proficiency into account, it could be concluded that especially the “amateur” group required objective criteria for the evaluation of a horse they are considering purchasing. Alongside “measureable” qualities, such as previous showing success or the level of training of the horse, also other attributes such as the simple handling of the horse should be taken into consideration. As particularly the “amateur” group in equestrian sport is increasing in numbers, it is therefore advisable when preparing a horse for sale to align oneself to the needs of this customer segment in order to ensure an effective and targeted marketing of horses.