Factors involved in the early pathogenesis of bovine Staphylococcus aureus mastitis with emphasis on bacterial adhesion and invasion. A review

Staphylococcus aureus is the most important and prevalent contagious mammary pathogen; it causes clinical and subclinical intramammary infection with serious economic loss and herd management problems in dairy cows. In vitro studies have shown that Staphylococcus aureus adheres to mammary epithelial cells and extracellular matrix components and invades into mammary epithelial as well as other mammary cells. Staphylococcus aureus strains from intramammary infection produce several cell surface-associated and extracellular secretory products. The exact pathogenic roles of most of the products and their effects on adhesion and invasion are not well evaluated. It is also known that mammary epithelial cell-associated molecules and extracellular matrix components interact with S. aureus during the pathogenesis of mastitis, but their roles on adhesion and invasion have not been characterized. The adhesion of S. aureus to epithelial cells may involve non-specific physicochemical interactions and/or specific interactions between bacterial cell-associated ligands and host cell surface receptors. In vitro adhesion depends on the S. aureus strain, the growth phase of the bacteria, the growth medium and the origin of the epithelial cells. Adhesion is hypothesized to be a prerequisite and crucial early step for mammary gland infection. Staphylococcus aureus invades mammary epithelial cells. It also invades other cells such as endothelial cells and fibroblasts. Bacteria are found enclosed in membrane bound vacuoles in the cytoplasm of mammary epithelial cells. Recent observations indicate that S. aureus escapes from the phagosome into the cytoplasm and induces apoptosis. The invasion into mammary epithelial cells may occur through an endocytic process that requires involvement of elements of the cytoskeleton or by direct binding of bacteria to epithelial cells through a process mediated by specific receptors that needs de novo protein synthesis by both cells. Thus, the recurrent subclinical infection may result from this intracellular existence of bacteria that are protected from host defenses and effects of antibiotics. This review emphasizes on recent findings on S. aureus adhesion to mammary epithelial cells and extracellular matrix components and invasion into mammary epithelial cells.     

Ultrasonographic anatomy and biometric analysis of the thoracic and abdominal organs in healthy foals from birth to age 6 months

Knowledge of normal renal parameters, as documented in mature horses, is essential for the accurate evaluation of abnormal kidneys. Although the ultrasonographic appearance and location of the abdominal organs in foals and the renal dimensions in neonates have been reported, there is currently no information available for the assessment of normal organ growth in foals. The objectives of the study were to describe the ultrasonographic characteristics, location and variations of the thoracic and abdominal organs with relation to age, height and weight; and provide a growth table for comparison with diseased foals. The thoracic and abdominal cavities of 10 healthy foals were evaluated at ages 1, 7, 14 and 21 days and 1, 2, 3, 4, 5 and 6 months. The equipment used was an Ausonics Opus Plus ultrasound. For every evaluation, weight and height were obtained, the foals were sedated and the area of study was clipped and cleaned. The ultrasonographic location, appearance and measurements of the different organs were recorded for each examination. The study revealed that foals age >1 month resemble the mature ultrasonographic pattern. Continual growth of the organs was observed from Day 1 to age 6 months, being faster in the first month. Organ growth was closely correlated with age, but not with sex, height or weight in healthy foals up to age 6 months. Our study has provided measurements of longitudinal organ growth in healthy foals, presented in a simple form for easy comparison with diseased individuals.     

In utero programming of sexually differentiated gonadotrophin releasing hormone (GnRH) secretion

It has long been recognised that steroids can have both organisational and activational effects on the reproductive neuroendocrine axis of many species, including the sheep. Specifically, if the ovine foetus is exposed to testosterone during a relatively short 'window' of in utero development (from approximately day 30-90 of a 147 day pregnancy) the neural mechanisms regulating gonadotrophin releasing hormone (GnRH) secretion become organised in a male-specific manner. In post-natal life the consequences of foetal androgen exposure are sexually differentiated responses of the GnRH neuronal network to activation by factors such as photoperiod and ovarian steroid hormones. Studies in the gonadectomized lamb have demonstrated that elevated concentrations of oestrogen (E) are unable to trigger a preovulatory-like GnRH surge in the male and the androgenized ewe lamb. Further, these animals have markedly reduced sensitivity to the inhibitory actions of progesterone on tonic GnRH release compared with normal ewes. The reasons for these abnormal steroid feedback mechanisms may reside in sexually dimorphic inputs to the GnRH neurone, including those from oestrogen-receptive neurones in the arcuate nucleus that synthetize the neuropeptide, neurokinin B (NKB). The consequences of in utero androgen exposure are reflected in a progressive and dramatic impairment of fertility in the ovary-intact ewe.     

Risk factors for stillbirths in two swine farms in the south of Brazil

We evaluated stillbirth risk factors in two commercial swine farms of the Rio Grande do Sul State (south of Brazil). The study was conducted during 1 month in Farm A and during 2 months in Farm B, both during 1999. Data for all farrowings that occurred during the study period were recorded (101 for Farm A and 373 for Farm B), without interference in the farm management. In Farm A, 39% of all litters born during the period of interest had stillborn piglets and the stillborn risk for piglets was 12%. In Farm B, 25% of all litters had stillborn piglets whereas the stillborn risk was 2%. Variables considered as potential risk factors for stillbirths were: parity (1, 2–3, 4+); breed (purebred or crossbred); sow body-condition (normal or fat); use of oxytocin during parturition (yes or no); obstetric intervention through vaginal palpation (yes or no); farrowing duration (<4 or ≥4 h); mummified fetuses (yes or no); total litter size (<12 or ≥12 piglets); and litter birth weight (<11 or ≥11 kg). All stillborn piglets had their classification validated by necropsy. In multivariable logistic-regressions, the cases were the litters having at least one stillborn piglet. In Farm A, litters having at least 12 pigs and in which oxytocin was used during the parturition had 20.8-times-higher odds of stillborn occurrence. In Farm B, litters from sows having parity ≥4 had 2.2-times-higher odds of stillborn occurrence than litters from parity 2 to 3 females, litters having ≥12 pigs had 2.0-times-higher odds of a stillborn piglet than smaller litters and farrowings in which vaginal palpation was performed had 8.0-times-higher odds. Farrowing room management to minimize stillborn risk should target higher-parity females, large litters and optimization of practices of obstetric interventions.     

Pheromone-Related Inhibitors of Ustilago hordei Mating and Tilletia tritici Teliospore Germination

ABSTRACT Ustilago hordei, the causal agent of barley covered smut, produces mating pheromones that break down to smaller peptide compounds that act as potent inhibitors of mating and germination in several fungi. The pheromones are members of the farnesylated family of proteins. Synthetic peptide analogs of the pheromone derivatives, ranging in size from 4 mers to full length pheromones, were farnesylated, methyl esterified, or both and tested for mating or teliospore germination inhibition with U. hordei or Tilletia tritici, respectively. N-Acetyl-S-farnesylcysteine, which inhibits processing of Ras, and other sulfur-containing compounds such as homocysteine or methionine, were likewise modified and tested. The most potent inhibitors were methionine methyl ester and modified 4-mer peptides from both pheromones. Alanine scanning of the inhibitory 4 mers determined that the native amino acid sequence was specific for a high level of activity. The sulfur amino acids appear to be required for inhibition. Glasshouse studies using selected antagonists of mating and teliospore germination as seed treatments inhibited covered smut of barley and common bunt of wheat, although the level of control was inconsistent. The use of pheromone-related antagonists to mating or teliospore germination is a promising, novel strategy for control of smut and bunt diseases.     

Molecular epidemiology of feline bordetellosis in two animal shelters in California,USA

"Kennel cough" in dogs in animal shelters is readily transmissible, reduces adoption rates, and commonly leads to the euthanasia of affected dogs. In cats, tracheobronchitis, conjunctivitis, and pneumonia have been associated with Bordetella bronchiseptica infection-but most cases of upper-respiratory infection (URI) probably are caused by herpesvirus and calicivirus, and many B. bronchiseptica culture-positive cats are clinically normal. Our prospective observational study was undertaken to document the contribution of B. bronchiseptica to disease in cats and dogs from two animal shelters undergoing outbreaks of canine kennel cough, to evaluate whether cross-species transmission might have occurred, and to determine if the presence of infected cats represented a risk to dogs. Clinically defined cases of kennel cough in dogs and URI in cats were investigated in two shelters by calculating clinical-disease incidence, alveolar-lavage cytological examination, bacterial and viral cultures, antibiotic-susceptibility testing, and molecular fingerprinting by pulsed-field gel electrophoresis.In a 40-cat and 40-dog "no-kill" shelter, the prevalences of culture positivity were 47% for B. bronchiseptica and 36% for calicivirus at the same time as two resident dogs demonstrated clinical cough. When no dogs had kennel cough 3 months later, 10% of cats were B. bronchiseptica-culture-positive and 63% calicivirus positive. In a large traditional shelter, the incidence of kennel cough in dogs increased over 12 weeks to a maximum of 19 cases/week/120 dogs, during which time the culture prevalence was 23% for B. bronchiseptica in dogs and 47% in cats. Three to 6 months before the kennel-cough epidemic, no dogs or cats were B. bronchiseptica positive. Very little genetic variability was detected in isolates from these shelters; all isolates except one corresponded to a single strain type which was identical to the pattern in a vaccine used in these shelters. Isolates from other cats, a horse, a llama, and a sea otter were genetically distinct from the shelter isolates. There was widespread resistance to cephalosporins and ampicillin, but low or no resistance to amoxicillin/clavulanate, trimethoprim-sulfamethoxazole, tetracycline, and enrofloxacin. Greater percent resistance was observed in the traditional shelter than in the no-kill shelter and feline isolates were more likely to be resistant than canine isolates.     

Reference values for activated coagulation time in cats

OBJECTIVE: To establish reference values for activated coagulation time (ACT) in cats by use of jugular venipuncture and direct collection of blood into ACT vacuum tubes. ANIMALS: 100 clinically normal cats that were to have elective surgery performed at a private practice. PROCEDURE: Collection of 3 blood samples for ACT measurement was attempted for each cat at the time of elective surgery: sample 1, obtained before sedation; sample 2, tube 1 of 2 consecutive samples obtained from a single venipuncture of the contralateral jugular vein after sedation with acepromazine and ketamine hydrochloride; and sample 3, tube 2 collected immediately following collection of sample 2 without removing the needle from the vein. Venipuncture quality was rated subjectively on a 3-point scale. RESULTS: Median ACT were 95 seconds for each sample group. The middle 95% of values ranged inclusively from 55 to 185 seconds (sample 1), 65 to 135 seconds (sample 2), 45 to 145 seconds (sample 3), and 55 to 165 seconds overall (samples 1, 2, and 3). Significant differences in ACT values were not detected between sample groups. Significant relationships between ACT and venipuncture quality or sex of cat were not detected. CONCLUSIONS AND CLINICAL RELEVANCE: With the ACT protocols used, clinically normal cats had ACT of < 165 seconds. The ACT in cats does not appear to be significantly affected by sex, sedation with acepromazine and ketamine, or by moderately traumatic venipunctures. These results refute widespread statements that ACT should be < 65 seconds in healthy cats. Cats with ACT repeatedly > 165 seconds should be further evaluated for hemostatic disorders.     

Comparison of the trotting gaits of Labrador Retrievers and Greyhounds

OBJECTIVE: To compare the trotting gaits of Labrador Retrievers and Greyhounds to determine whether differences in locomotion are attributable to differences in their manner of moving or to body size and shape differences between these 2 breeds. ANIMALS: 8 healthy 5-month-old Greyhounds and 5 healthy Labrador Retrievers between 6 and 18 months old. PROCEDURE: A series of 4 force platforms was used to record independent ground reaction forces on the forelimbs and hind limbs during trotting. Values of stride parameters were compared between breeds before and after normalization for size differences. Standard values of absolute and normalized stride period and stride length were determined from linear regressions of these parameters on relative (normalized) velocity. Forces were normalized to body weight and compared at the same relative velocity. RESULTS: Greyhounds used fewer, longer strides than the Labrador Retrievers to travel at the same absolute speed. After normalization for body size differences, most measurable differences between breeds were eliminated. Subtle differences that did persist related to proportion of the stride that the forefoot was in contact with the ground, timing of initial hind foot contact relative to initial forefoot contact, and distribution of vertical force between the forelimbs and hind limbs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that apparent differences in the trotting gait between Labrador Retrievers and Greyhounds are mainly attributable to differences in size, and that dogs of these 2 breeds move in a dynamically similar manner at the trot.