Occurrence of fruiting structures allows determination of Purpureocillium lilacinum as an inciting agent of pleuritis and pneumonia in a loggerhead sea turtle (Caretta caretta) by histopathologic correlation to culture.

Purpureocillium lilacinum and Beauveria bassiana were isolated from lung sampled at necropsy of a 12 year-old female loggerhead sea turtle (Caretta caretta) that had displayed abnormal buoyancy. Histopathologic evaluation revealed pleuritis and pneumonia with non-melanized, septate hyphae and fruiting structures identical to those of P. lilacinum. This case emphasizes the importance of a histological correlate to fungal culture when environmental fungi are isolated and demonstrates the infrequent phenomenon of fruiting or conidial production in tissue.

Evaluation of a clay-based acidic bedding conditioner for dairy cattle bedding.

This study investigated the effects of a clay-based acidic bedding conditioner on sawdust bedding pH, dry matter (DM), environmental pathogen counts, and environmental bacterial counts on teat ends of lactating dairy cows. Sixteen lactating Holstein cows were paired based on parity, days in milk, milk yield, and milk somatic cell count, and were negative for the presence of an intramammary pathogen. Within each pair, cows were randomly assigned to 1 of 2 treatments with 3-wk periods in a crossover design. Treatment groups consisted of 9 freestalls per group bedded with either untreated sawdust or sawdust with a clay-based acidic bedding conditioner, added at 3- to 4-d intervals over each 21-d period. Bedding and teat ends were aseptically sampled on d 0, 1, 2, 7, 14, and 21 for determination of environmental bacterial counts. At the same time points, bedding was sampled for DM and pH determination. The bacteria identified in the bedding material were total gram-negative bacteria, Streptococcus spp., and coliform bacteria. The bacteria identified on the teat ends were Streptococcus spp., coliform bacteria, and Klebsiella spp. Teat end score, milk somatic cell count, and intramammary pathogen presence were measured weekly. Bedding and teat cleanliness, environmental high and low temperatures, and dew point data were collected daily. The bedding conditioner reduced the pH, but not the DM, of the sawdust bedding compared with untreated sawdust. Overall environmental bacterial counts in bedding were lower for treated sawdust. Total bacterial counts in bedding and on teat ends increased with time over both periods. Compared with untreated sawdust, the treated bedding had lower counts of total gram-negative bacteria and streptococci, but not coliform counts. Teat end bacterial counts were lower for cows bedded on treated sawdust for streptococci, coliforms, and Klebsiella spp. compared with cows bedded on untreated sawdust. The clay-based acidic bedding conditioner reduced environmental pathogens in sawdust bedding and teat ends without affecting teat end integrity.

Antibacterial effect of plant-derived antimicrobials on major bacterial mastitis pathogens in vitro.

The objective of this study was to investigate the antimicrobial effect of plant-derived antimicrobials including trans-cinnamaldehyde (TC), eugenol, carvacrol, and thymol on major bacterial mastitis pathogens in milk. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the aforementioned compounds on Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Staphylococcus aureus, and Escherichia coli were determined. In addition, the bactericidal kinetics of TC on the aforementioned pathogens and the persistence of the antimicrobial activity of TC in milk over a period of 2 wk were investigated. All 4 plant-derived molecules exhibited antimicrobial activity against the 5 mastitis pathogens tested, but TC was most effective in killing the bacteria. The MIC and MBC of TC on Staph. aureus, E. coli, and Strep. uberis were 0.1 and 0.45%, respectively, whereas that on Strep. agalactiae and Strep. dysgalactiae were 0.05 and 0.4%, respectively. The MIC and MBC of the other 3 molecules ranged from 0.4 to 0.8% and 0.8 to 1.5%, respectively. In time-kill assays, TC at the MBC reduced the bacterial pathogens in milk by 4.0 to 5.0 log(10) cfu/mL and to undetectable levels within 12 and 24 h, respectively. The antimicrobial effect of TC persisted for the duration of the experiment (14 d) without any loss of activity. Results of this study suggest that TC has the potential to be evaluated as an alternative or adjunct to antibiotics as intramammary infusion to treat bovine mastitis.

Fatal necrotising fasciitis and myositis in a cat associated with Streptococcus canis.

Necrotising fasciitis and necrotising myositis are rare but serious life threatening conditions reported mainly in human beings and dogs. Most cases to date have been caused by beta-haemolytic streptococci of Lancefield groups A, B, C or G. Necrotising fasciitis has been reported only twice in cats and necrotising myositis has never been described. This paper describes a fatal case of necrotising fasciitis and myositis with pneumonia and septicaemia in a nine-year-old cat. The cat had been undergoing treatment for a suspected tear of the cranial cruciate ligament, but on the seventh day of treatment it suddenly deteriorated and died. On postmortem examination, there was an area of hair loss from its left hindlimb and discoloration of the underlying fascia and biceps femoris muscle. Severe necrotising fasciitis and myositis, with numerous intralesional Gram-positive coccoid bacteria, was diagnosed histologically. Other findings included necrotising pneumonia, pleuritis, focal encephalitis, myocarditis and nephritis. Culture of the affected tissues yielded a pure, heavy growth of Streptococcus canis.

Extraintestinal pathogenic Escherichia coli-induced acute necrotizing pneumonia in cats.

Extraintestinal pathogenic Escherichia coli (ExPEC) are pathogens involved in several disease conditions, ranging from urinary tract infection to meningitis in humans and animals. They comprise epidemiologically and phylogenetically distinct strains, affecting most species and involving any organ or anatomical site. Here, we report fatal cases of necrotizing pneumonia in cats. Over a 1-week period, 13 cats from an animal shelter in Stamford, Connecticut were presented for necropsy. All had a clinical history of acute respiratory disease. The gross and microscopic findings for all the cats were consistent. Escherichia coli was uniformly isolated from the lungs of all the tested cats. All the isolates were haemolytic, genetically related as determined by enterobacterial repetitive intergenic consensus PCR, and harboured genes encoding for cytotoxic necrotizing factor-1 and fimbriae and adhesions that are characteristic of ExPEC, implying a point source clonal outbreak. As cats are common household pets, this report raises concerns regarding zoonotic potential (in either direction) for these ExPEC strains.

Antibacterial effect of caprylic acid and monocaprylin on major bacterial mastitis pathogens.

Bovine mastitis is the most significant economic drain on the worldwide dairy industry. Concerns regarding poor cure rates, emergence of bacterial resistance, and residues in milk necessitate development of alternative therapeutic approaches to antibiotics for treatment of mastitis. A variety of free fatty acids and their monoglycerides have been reported to exert antimicrobial activity against a wide range of microorganisms. The objective of our study was to examine the efficacy of caprylic acid, a short-chain fatty acid, and its monoglyceride, monocaprylin, to inactivate common mastitis pathogens, including Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Staphylococcus aureus, and Escherichia coli. Milk samples containing 50 mM or 100 mM caprylic acid, and 25 mM or 50 mM monocaprylin were inoculated separately with a 3-isolate mixture of each of the 5 pathogens, and incubated at 39 degrees C. Populations of surviving bacteria were determined at 0 min, 1 min, 6 h, 12 h, and 24 h of incubation. Both caprylic acid and monocaprylin reduced all 5 pathogens by >5.0 log cfu/mL after 6 h of incubation. Among the bacterial species tested, Strep. agalactiae, Strep. dysgalactiae, and Strep. uberis were most sensitive, and E. coli was most tolerant to caprylic acid and monocaprylin. Results of this study indicate that caprylic acid and monocaprylin should be evaluated as alternatives or adjuncts to antibiotics as intra-mammary infusion to treat bovine mastitis.

Mycobacterium marinum dermatitis and panniculitis with chronic pleuritis in a captive white whale (Delphinapterus leucas) with aortic rupture.

A 16-year-old female white whale, Delphinapterus leucas, died after nearly 18 months of chronic lymphopenia and pyogranulomatous dermatitis. Necropsy revealed rupture of the aorta with hemorrhage into the cranial mediastinum and between fascial planes of the ventral neck musculature. Multiple foci of ulcerative dermatitis and panniculitis were present across the thorax and abdomen and surrounded the genital folds. In addition, there was a chronic proliferative pleuritis with over 20 liters of histiocytic exudate in the thoracic cavity. Acid-fast bacteria consistent with Mycobacterium sp. were identified in sections of skin lesions and in cytospins of pleural exudate. Cultures of pleura and 1 skin lesion collected at necropsy yielded sparse growth of an acid-fast bacillus with colony characteristics and morphology consistent with Mycobacterium marinum. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis confirmed the presence of M. marinum DNA in samples of skin. This is the first documented occurrence of mycobacteriosis in a white whale and is a unique presentation of mycobacterial dermatitis and panniculitis with chronic pleuritis in a cetacean. The improved PCR-RFLP protocol utilized in this case unifies techniques from several protocols to differentiate between species of Nocardia and rapidly growing mycobacteria clinically relevant to aquatic animals.

Masticator space abnormalities associated with mandibular osteoradionecrosis: MR and CT findings in five patients.

BACKGROUND AND PURPOSE: Imaging of patients with a clinical diagnosis of mandibular osteoradionecrosis (ORN) is often performed to support that clinical suspicion, evaluate the extent of the disease, or exclude coexistent tumor recurrence. The purpose of our study was to describe the clinical, MR imaging, and CT features of five patients with mandibular ORN associated with prominent soft-tissue abnormality in the adjacent masticator muscles. METHODS: The MR and CT examinations of five patients with mandibular ORN associated with soft-tissue abnormalities in the adjacent masticator muscles were reviewed. All patients had received external beam radiotherapy for primary head and neck malignancies, with a total radiation dose range of 60 Gy to 69 Gy in 30 to 38 fractions. RESULTS: CT revealed the typical osseous findings of cortical disruption, trabecular disorganization, and fragmentation in all five patients. Abnormal diffuse enhancement of the adjacent masseter and pterygoid muscles was noted in all patients. Four patients had prominent mass-like thickening of these muscles adjacent to the osseous abnormality. Of the three patients who underwent MR imaging, all showed homogeneous abnormal T1 hypointensity, T2 hyperintensity, and intense enhancement of the bone marrow in the involved mandible. The masticator muscles adjacent to the osseous abnormality also showed abnormal T2 hyperintensity and intense diffuse enhancement on MR images. CONCLUSION: Mandibular ORN can be associated with prominent soft-tissue thickening and enhancement in the adjacent musculature. These changes can appear mass-like and are not related to tumor recurrence or metastatic disease.

Inducible and constitutive in vitro neutrophil chemokine expression by mammary epithelial and myoepithelial cells.

Previously, our laboratory showed that bovine and caprine mammary secretions are chemotactic and that chemoattractants found in these secretions are qualitatively different according to infection status and/or lactation stage. However, the cellular source of the chemoattractants has not been defined. In this study we used a modified Boyden chamber assay to examine the ability of previously established caprine mammary epithelial cell (CMEC) and myoepithelial cell (CMMyoEC) lines to produce chemoattractants for neutrophils. We found that CMEC culture supernatants, but not those of CMMyoEC cultures, induced in vitro neutrophil chemotaxis. Further characterization showed that chemotactic activity was produced when the cells underwent contact-induced differentiation. Neutrophil migration was chemotactic, not chemokinetic, and was augmented when the epithelial and myoepithelial cells were cocultured. Additionally, chemotactic activity was inducible by Staphylococcus aureus plus alpha-toxin, Escherichia coli, and interleukin-1beta (IL-1beta) in CMEC cultures. However, CMMyoEC cultures could not be induced to produce chemotactic activity. Anti-IL-8 antibody was able to block some constitutively produced chemotactic activity and chemotactic activity induced by IL-1beta and S. aureus plus alpha-toxin. These results indicate that epithelial cells may play a major role in producing chemoattractants, specifically IL-8, in the mammary gland.

Oral cisapride for the control of delayed vomiting following high-dose cisplatin.

Although combination antiemetics prevent vomiting during the initial 24 h after high-dose (> or =100 mg/m2) cisplatin, many patients experience delayed emesis 24-120 h afterwards despite receiving prophylactic dexamethasone and metoclopramide during this time. Cisapride is a prokinetic agent, which stimulates propulsive motility throughout the gastrointestinal tract without causing extrapyramidal effects. In this phase II trial, we tested the ability of cisapride to prevent delayed emesis following cisplatin. Twenty patients receiving initial cisplatin >100 mg/m2 were entered. All patients received intravenous dexamethasone with either metoclopramide or ondansetron to prevent acute emesis 0-24 h after receiving cisplatin. Patients who had experienced two or fewer acute vomiting episodes then received cisapride 20 mg orally four times daily for 4 days (24-120 h after cisplatin). Cisapride prevented delayed emesis in 2 patients (10%) during the entire 4-day period (95% confidence interval, 1-32%). Abdominal cramping and pain occurred in 35%. At the dose and schedule tested, oral cisapride prevented delayed emesis in only 10% of patients receiving cisplatin >100 mg/m2 and caused abdominal cramping in 35%. Since in prior trials among similar patients, placebo prevented delayed emesis in 11%, further study of cisapride and dose escalation for this indication are not recommended.

Physiologic-chemoattractant-induced migration of polymorphonuclear leukocytes in milk.

The somatic cell count (SCC; leukocytes and epithelial cells) in milk is used as an indicator of udder health status. A SCC above the regulatory standard is generally considered as an indication of mastitis. Therefore, milk with a SCC equal to or greater than the regulatory limit cannot be sold to the public because it is unsuitable for human consumption. This study was performed to determine whether SCC levels above the regulatory limit observed in goats during late lactation are a physiologic or a pathological response of the goat mammary gland. Differential counts of cells in nonmastitic goat milk samples during late lactation revealed that approximately 80% of the cells were polymorphonuclear leukocytes (PMNs). In addition, microchemotaxis assay results indicated that normal nonmastitic late-lactation-stage goat milk is significantly higher (P < 0.001) in PMN chemotactic activity than early-lactation-stage goat milk, with a mean chemotactic activity of 14.9 and 42.7/mg of protein for early and late lactation stages, respectively. Physicochemical analyses also suggest that the PMN infiltration observed in normal late-lactation-stage goat milk is due to a PMN chemotactic factor(s) that is different from the PMN chemotactic factor(s) present in mastitic milk. Interestingly, the PMN chemotactic factor in late-lactation-stage goat milk is highly acid resistant (pH 2), suggesting that the factor is able to survive the highly acidic gastric environment and may therefore be important in the augmentation of the immune systems of sucklings. These results indicate that the chemotactic factor(s) present in the milk of normal late-lactation-stage goats is nonpathological and may play a physiologic regulatory role in mammary gland involution. Hence, the regulatory standard for goat milk needs to be redefined in order to reflect this.

Mycoplasmal conjunctivitis in a European starling.

Bilateral conjunctivitis and episcleritis were identified in an adult European starling (Sturnus vulgaris). A novel mycoplasma species, Mycoplasma sturni, was isolated in pure culture from the conjunctiva of both eyes. The clinical presentation was similar to that of conjunctivitis in house finches (Carpodacus mexicanus) caused by Mycoplasma gallisepticum. However, the histologic lesions were distinct, by the presence of ulceration and by the absence of epithelial hyperplasia and lymphoplasmacytic infiltration [corrected]. Mycoplasma sturni ferments glucose, does not hemadsorb or hemagglutinate chicken erythrocytes, and grows rapidly at 37 C in comparison to other Mycoplasma spp. The role of M. sturni in conjunctivitis in other passerine species is presently unknown.

Mycoplasma sturni sp. nov., from the conjunctiva of a European starling (Sturnus vulgaris).

Strain UCMF(T) (T = type strain) was isolated from the conjunctiva of a European starling (Sturnus vulgaris) with conjunctivitis. Colonies grown on conventional mycoplasma agar possessed the typical fried-egg appearance observed with many mycoplasmal species. Electron micrographs of ultrathin sections of UCMF(T) revealed a pleomorphic cellular morphology; the cells ranged from spherical to elliptical or flask shaped. The cell size ranged from 0.3 to 0.5 microns. Strain UCMF(T) grows well in a variety of mycoplasma broth formulations at 25 degrees C, with rapid and heavy growth at 37 degrees C. No growth occurs at 42 degrees C. This organism ferments glucose but does not hydrolyze urea or arginine and has an absolute requirement for sterol from growth. Strain UCMF(T) does not hemagglutinate or hemadsorb chicken erythrocytes. The genome size is 870 kbp, and the guanine-plus-cytosine content is 31 mol%. Sequence analysis of the 16S rRNA gene demonstrated that this organism is unique and has not been described previously. Serological analysis confirmed that strain UCMF(T) is distinct from all previously identified Mycoplasma, Acholeplasma, Spiroplasma, Entomoplasma, and Mesoplasma species. This organism represents a new species, for which we propose the name Mycoplasma sturni. Strain UCMF (= ATCC 51945) is the type strain of M. sturni sp. nov.

Antibiotic susceptibility profiles for mastitis treatment.

Susceptibility tests were performed on milk samples representing prevalent mastitis infections in certain herds. Susceptibility patterns of the same bacterial species from several mastitis infections in the same herd were consistent. The herd antibiotic susceptibility profiles were used as a basis for selecting antibiotics for treatment of all such mastitis cases in that herd. A high degree of correlation was seen between the susceptibility test results and treatment results. Susceptibility patterns of the same bacterial species from mastitis infections in different herds varied greatly, which indicated that any one antibiotic would not work equally well against the same bacterial infection in every herd. Therefore, treatment should be selected on the basis of susceptibility test results. When both Streptococcus and Staphylococcus mastitis occurred in the same herd, the susceptibility patterns for the 2 bacterial species varied widely. Therefore, for herds that experienced both streptococcal and staphylococcal mastitis, antibiotics to which both bacterial species were susceptible were used for treatment.