Experimental studies on the impact of an increased dose of anionic salts on the metabolism of dairy cows

Summary

This study was initiated to investigate the influence of a daily dose of anionic salts (AS) above the valid upper limit at present on metabolism of dairy cows. Eleven non‐pregnant and non‐lactating Holstein‐Friesian crossbred cows with a permanent rumen cannula were used in a study with a controlled feeding design. The initial daily dose was 2500 meq/day, which resulted in a Dietary Cation Anion Difference (DCAD) of ‐211 meq/kg dry matter. Every seven days, the daily dose was raised by 500 meq. If a cow stopped eating, the application of AS was stopped and these cows were monitored over the next seven days. On day 30 another batch of hay, having the same DCAD but higher concentrations of minerals and energy, was fed. Blood and urine samples were taken to monitor acid‐base balance and calcium concentrations. Acid‐base balance was strongly influenced by AS. Blood pH dropped steadily and reached values around 7.23. Urine pH dropped quickly below 6 and remained at that level regardless of the increased dosage of AS. Net acid base excretion (NABE) fell continuously with the increase of the dosage of AS and reached values below ‐200 mmol/l. Calcium concentrations in the serum were nearly stable, but those in urine increased sharply and remained on an elevated level with increasing doses of AS. A few days before the individual cow's refusal of feed intake, calcium excretion in urine decreased. The majority of cows stopped eating while consuming a diet containing 3500 to 4000 meq AS except two animals who consumed up to 6000 meq/day AS but they received the better hay in the second half of the treatment period. In this time pH in blood increased slowly. NABE remained stable on a low level at ‐100 mmol/l. The results showed that with an increasing amount of AS fed the risk of clinical acidosis increased. The decreasing urine concentrations of calcium indicate a breakdown of the compensation capability of the single cow. Besides the dose of AS fed, the quality of the feed stuff might be another factor concerning the tolerance of cows against AS.     

Morphometric analyses of the body and the branches of the normal third interosseous muscle (suspensory ligament) in Standardbreds

The third interosseous muscle (suspensory ligament, TIOM) is composed of connective tissue (CT) with a variable proportion of muscle (MT) and adipose tissue (AT). The aim of our study is to quantify the CT, MT and AT within the body and the branches of right thoracic and pelvic limbs TIOM in sound horses to determine whether there are differences in CT, MT and AT between age, sex, limbs and levels. Right limbs from 11 sound horses were collected. Samples from 6 levels of the TIOM were embedded in paraffin or in Tissue‐Tek^®. Most of the paraffin sections were shredded. Using the cryosection, some artefacts appeared. Cryoprotection was carried out, which produced the best results. Hematoxylin–phloxine–saffron and Hematoxylin–eosin gave a good contrast of colours between the tissues observed allowing the use of an image analysis programme to calculate percentage of each tissue within the TIOM. The percentage of MT and AT decreased significantly (P < 0.0001), whereas the percentage of CT increased significantly (P < 0.0001) with age and when descending from the proximal to the distal level of the TIOM. The percentage of MT was significantly higher (P < 0.0001) in females than males, while the percentage of CT was significantly higher (P < 0.0001) in males than females. The percentage of AT was significantly higher (P = 0.0278) in pelvic limbs than in thoracic limbs. These results confirm the variation in tissue composition within the TIOM of sound horses.     

VERTEBRAL HEART SCORES IN EIGHT DOG BREEDS

The vertebral heart score (VHS) measurement is commonly used to provide a more objective measurement of cardiomegaly in canines. However, several studies have shown significant breed variations from the value previously established by Buchanan and Bücheler (9.7 ± 0.5). This study describes VHS measurements in Pug, Pomeranian, Yorkshire Terrier, Dachshund, Bulldog, Shih Tzu, Lhasa Apso, and Boston Terrier dog breeds. Dogs with two or three view thoracic radiographs, no subjective radiographic evidence of cardiomegaly, and no physical examination findings of heart murmurs or gallop rhythms were included in the study. The Pug, Pomeranian, Bulldog, and Boston Terrier groups were found to have a VHS significantly greater than 9.7 ± 0.5 (P < 0.00001, P = 0.0014, P < 0.0001, P < 0.00001, respectively). Body condition score (BCS) was found to have a significant effect on the VHS of Lhasa Apso group. Anomalous vertebrae in the thoracic column were associated with a significant increase in VHS of the Bulldog (P = 0.028) and Boston Terrier (P = 0.0004) groups. Thoracic depth to width ratio did not have a significant effect on VHS.     

Distribution and Genotypic Characterization of Campylobacter jejuni Isolated from Poultry in Split and Dalmatia County,Croatia

Consumption of poultry contaminated with Campylobacter jejuni has been recognized worldwide as the leading cause of campylobacteriosis. Therefore, the aim of our study was to investigate the prevalence and genotype diversity of Campylobacter jejuni in poultry meat intended for consumption in Split and Dalmatia County, which is the second biggest County in Croatia. Furthermore, we also wanted to discover possibly stable clones of C. jejuni appearing in different samples and periods of time, which would indicate their ability to persist in or adapt to poultry. In the period from March 2008 until June 2010, 834 samples of poultry from various sources were examined using a surface swab technique. Isolation of C. jejuni was performed by Preston broth and Karmali agar. Identification of the isolates was carried out using biochemical tests. C. jejuni was found in 84 of 574 chicken samples (14.6%) and in nine of 260 samples of turkey (3.5%). Pulse‐field gel electrophoresis (PFGE) was used to analyse 61 obtained isolates using SmaI and KpnI. Of 22 different macrorestriction profiles (MRP) that were found, five were detected in poultry from both different locations and periods of time. Samples from 11 locations were found to be contaminated with more than two different genotypes of C. jejuni. Interestingly, the same MRP were found both in poultry declared to be of domestic origin and in the poultry imported from abroad. The prevalence of C. jejuni in poultry samples was in accordance with previously reported results. Genotypic analysis indicated that the population of C. jejuni in Split and Dalmatia County was diverse and that multiple strains of C. jejuni could be found in the same poultry samples. Furthermore, the same genotypes were identified from the samples obtained from different locations and periods of time, which could support the theory of a global existence of certain MRP that are able to persist in or adapt to poultry.     

Effect of injecting amino acids into the egg on embryonic and subsequent growth in the domestic fowl

1. The growing embryo uses about 1% of the total protein available in the egg during the first week of incubation.

2. Injecting the growing embryo with amino acids equivalent to the amount of protein used resulted in higher weight of chicks at all stages up to 56 d.     

MAGNETIC RESONANCE IMAGING OF A MEDULLARY DERMOID CYST WITH SECONDARY HYDROCEPHALUS IN A DOG

Magnetic resonance images were acquired of the brain of a 7-year-old male Golden Retriever with hydrocephalus secondary to a medullary lesion. Images were acquired prior to and 4 weeks following surgical treatment for the hydrocephalus, and the dog was euthanased following the second imaging session. The MR images demonstrated a medullary lesion with patchy but predominantly hyperintense signal with both T1- and T2-weighting, within which small areas of low signal were scattered. There was little edema associated with this lesion and no enhancement with gadolinium. Postmortem examination revealed the medullary mass to be a dermoid cyst. Several small nodular lesions were identified within the central nervous system on the magnetic resonance images whose origin was uncertain on postmortem examination.     

Aneurysmal dilatation of the right auricle in two dogs

An 8-year-old sexually intact male Golden Retriever with a history of collapse during exercise underwent an examination during which tachydysrhythmia was identified. At another institution, a 12.5-year-old spayed female Lhasa Apso was referred because of a cough and for evaluation of a heart murmur. In the Golden Retriever, radiographic examination revealed bulging of the craniodorsal aspect of the cardiac silhouette and echocardiography revealed right atrial dilatation. In the Lhasa Apso, a cranial mediastinal mass was suspected on the basis of radiographic findings, but no abnormalities were detected echocardiographically. In both dogs, nonselective angiography and contrast-enhanced computed tomography revealed a markedly enlarged, thin-walled right auricle. Exploratory thoracotomy in the Golden Retriever revealed a defect in the pericardium through which the right auricle and part of the atrium had herniated. In dogs, a right auricular aneurysm should be considered in differential diagnoses of a cranial mediastinal mass (detected radiographically) adjacent to the cardiac silhouette.     

Culture of Differentiated Adult Rabbit Auricular Chondrocytes

Chondrocytes dedifferentiate to a fibroblast‐like phenotype on plastic surfaces. Dedifferentiation is reversible if these cells are then cultured embedded in gels as alginate, agarose or collagen. Chondrocytes cultured in suspension on a non‐adherent surface are also known to form aggregates of differentiated cells. The knowledge of chondrocyte behavior in culture is relevant for tissue engineering purposes. In this report we describe a simple method to culture differentiated or redifferentiated rabbit auricular chondrocytes on plastic surfaces with a stable phenotype. When chondrocyte aggregates formed in suspension are next seeded on plastic surfaces, most of them attach to the plastic as round or polygonal cells, and this morphological differentiation, confirmed by the presence of type II collagen, is stable for long culture periods. We also report that the addition of aggregates to monolayer cultures of dedifferentiated chondrocytes results in their redifferentiation, as is shown by their morphological changes and the synthesis of type II collagen. Therefore, this simple method can be useful for the study of chondrocyte behavior on plastic surfaces and for redifferentiating previously proliferated chondrocytes in tissue engineering techniques. Furthermore, these results demonstrate that, in addition to culture conditions such as cell isolation method or cell‐density, chondrocyte behavior on plastic depends on the presence or absence of aggregates resulting from the dissociation process.     

Comparative study of biochemical methods for the identification of Dichelobacter nodosus

The aim of this study was to compare four identification procedures to detect Dichelobacter nodosus and develop a rapid, simple and effective method to identify D. nodosus strains isolated from cases of ovine footrot. The four methods used were: (a) the classic guidelines set down by Holdeman et al. (1977) and Summanen et al. (1993) which are based on gas liquid chromatography (GLC) and different biochemical tests, this method was considered as landmark; (b) Baron and Citron's flowchart for the rapid identification of Gram-negative rod-shaped anaerobes (1997); (c) the API rapid 32 A system (bio Mérieux), and (d) Mast ID Anaerobe ID Ring (MID8) (Mast Diagnostics). None of the four methods used allowed us to correctly identify the D. nodosus strains (neither the strains isolated from cases of ovine footrot nor those originating from type collection). Because of the difficulties encountered in obtaining a correct identification of D. nodosus, we propose a simple, rapid and effective way to achieve this task. Our flowchart will provide the means to identify this microorganism in any laboratory of general microbiology without having to use any specialised equipment.