The effect of transportation and lairage on faecal shedding and carcass contamination with Escherichia coli O157 and O26 in very young calves in New Zealand.

The effect of transportation and lairage on the faecal shedding and post-slaughter contamination of carcasses with Escherichia coli O157 and O26 in young calves (4-7-day-old) was assessed in a cohort study at a regional calf-processing plant in the North Island of New Zealand, following 60 calves as cohorts from six dairy farms to slaughter. Multiple samples from each animal at pre-slaughter (recto-anal mucosal swab) and carcass at post-slaughter (sponge swab) were collected and screened using real-time PCR and culture isolation methods for the presence of E. coli O157 and O26 (Shiga toxin-producing E. coli (STEC) and non-STEC). Genotype analysis of E. coli O157 and O26 isolates provided little evidence of faecal-oral transmission of infection between calves during transportation and lairage. Increased cross-contamination of hides and carcasses with E. coli O157 and O26 between co-transported calves was confirmed at pre-hide removal and post-evisceration stages but not at pre-boning (at the end of dressing prior to chilling), indicating that good hygiene practices and application of an approved intervention effectively controlled carcass contamination. This study was the first of its kind to assess the impact of transportation and lairage on the faecal carriage and post-harvest contamination of carcasses with E. coli O157 and O26 in very young calves.

Nationwide prevalence and risk factors for faecal carriage of Escherichia coli O157 and O26 in very young calves and adult cattle at slaughter in New Zealand.

Nationwide prevalence and risk factors for faecal carriage of Escherichia coli O157 and O26 in cattle were assessed in a 2-year cross-sectional study at four large slaughter plants in New Zealand. Recto-anal mucosal swab samples from a total of 695 young (aged 4-7 days) calves and 895 adult cattle were collected post-slaughter and screened with real-time polymerase chain reaction (PCR) for the presence of E. coli O157 and O26 [Shiga toxin-producing E. coli (STEC) and non-STEC]. Co-infection with either serogroup of E. coli (O157 or O26) was identified as a risk factor in both calves and adult cattle for being tested real-time PCR-positive for E. coli O157 or O26. As confirmed by culture isolation and molecular analysis, the overall prevalence of STEC (STEC O157 and STEC O26 combined) was significantly higher in calves [6·0% (42/695), 95% confidence interval (CI) 4·4-8·1] than in adult cattle [1·8% (16/895), 95% CI 1·1-3·0] (P < 0·001). This study is the first of its kind in New Zealand to assess the relative importance of cattle as a reservoir of STEC O157 and O26 at a national level. Epidemiological data collected will be used in the development of a risk management strategy for STEC in New Zealand.

Diversity and relatedness of Shiga toxin-producing Escherichia coli and Campylobacter jejuni between farms in a dairy catchment.

The aim of this study was to examine the population structure, transmission and spatial relationship between genotypes of Shiga toxin-producing Escherichia coli (STEC) and Campylobacter jejuni, on 20 dairy farms in a defined catchment. Pooled faecal samples (n = 72) obtained from 288 calves were analysed by real-time polymerase chain reaction (rtPCR) for E. coli serotypes O26, O103, O111, O145 and O157. The number of samples positive for E. coli O26 (30/72) was high compared to E. coli O103 (7/72), O145 (3/72), O157 (2/72) and O111 (0/72). Eighteen E. coli O26 and 53 C. jejuni isolates were recovered from samples by bacterial culture. E. coli O26 and C. jejuni isolates were genotyped using pulsed-field gel electrophoresis and multilocus sequence typing, respectively. All E. coli O26 isolates could be divided into four clusters and the results indicated that E. coli O26 isolates recovered from calves on the same farm were more similar than isolates recovered from different farms in the catchment. There were 11 different sequence types of C. jejuni isolated from the cattle and 22 from water. An analysis of the population structure of C. jejuni isolated from cattle provided evidence of clustering of genotypes within farms, and among groups of farms separated by road boundaries.

Histomorphological and immunohistochemical findings in testes, bulbourethral glands and brain of immunologically castrated male piglets.

The aim of this study was the histological and immunohistochemical evaluation and comparison of testicular, bulbourethral and brain tissue in immunized and intact control boars. Fourteen male piglets, aged between 10 and 16 weeks, were vaccinated twice subcutaneously 4 to 5 weeks apart with Improvac, an anti-GnRH vaccine. The pigs were sacrificed 1 to 16 weeks following the second injection. Testicular weight was recorded and various tissue samples were collected and fixed in formalin and Bouin's fixative for histological examination. In addition, 2 boars were immunized five times and slaughtered 60 weeks after the last injection. Histological and immunohistological studies performed on testes and epididymes showed clear signs of atrophy in the immunized animals and a significant reduction in paired testes weight was seen in treated boars. Microscopically, the mean diameter of the seminiferous tubules was markedly reduced. Spermatogonia as well as few spermatocytes were visible between the Sertoli cells and Leydig cells were atrophic. None or only few spermatozoa were detected in the epididymis. The bulbourethral glands of immunocastrated pigs were smaller than in control pigs and showed histological evidence of atrophy. Immunohistological detection of LH and FSH in the pituitary gland of treated and control boars showed no quantifiable difference in the amount of these two gonadotropins and no lesions were visible in the hypothalamus and the pituitary gland. From our findings it can be concluded that the anti-GnRH vaccine Improvac induces severe atrophy of testes and bulbourethral glands in immunized pigs. This effect appears to be reversible, depending on the immune response of each animal and the time elapsed after the last booster injection.

Hemangiopericytoma of the ciliary body.

A 60-year-old woman had a ciliary body tumor that resembled an amelanotic malignant melanoma by clinical examination. The magnetic resonance imaging scan correlated with the clinical diagnosis. Because of apparent rapid growth, her eye was enucleated. However, histopathologic examination revealed a highly vascularized, spindle-cell tumor harboring immunoreactive actin and vimentin. Electron microscopy further supported the diagnosis of hemangiopericytoma with smooth-muscle differentiation. To our knowledge, this is the first report of a hemangiopericytoma of the ciliary body.

Pingueculae and pterygia.

Pingueculae and pterygia are benign peribulbar lesions composed of degenerated basophilic subepithelial tissue. Pingueculae do not affect vision, and minor irritation can usually be managed with artificial tears. Pterygia may affect the visual axis and require surgical and adjunctive treatment. The various therapeutic strategies are reviewed. A conservative approach is advocated, as surgical removal of primary pterygia may result in recurrent ptergyia that are more difficult to manage than the primary lesions.

Opioid receptor types for endogenous enkephalin in the thoracic ganglion of the crab, Carcinus maenas.

In crustaceans, the endogenous opioid peptides, enkephalins, are known to be concentrated in the thoracic ganglion, although they have been demonstrated in all parts of the nervous system. Bmax and Kd measurements have been obtained for the binding of ligands used to characterize delta- and kappa-type opioid receptors in vertebrates. High- and low affinity binding of [3H] [2-D-Pen5-D Pen] enkephalin ([3H]DPDPE) has been measured with a Kd = 9.2 +/- 2.4 nM, Bmax = 153 fmol/mg, and Kd = 243 +/- 27 nM, Bmax = 1.785 pmol/mg, respectively. In addition a kappa-type receptor with Kd 85.5 +/- 12.6 nM and Bmax = 21.138 pmol/mg protein has been recorded. Binding characteristics of several ligands were monitored. Electrophoretic studies of affinity chromatographically purified receptor fractions revealed a molecular mass of 60 kDa. Isoelectric focusing showed a specific binding of [3H]DPDPE to thoracic ganglion membranes at a pl of 5.5.

Delta and kappa opioid receptors in eyestalk ganglia of a crustacean.

Crustacean eyestalk ganglia are part of the protocerebrum and have been demonstrated to produce numerous neurohormones. 3H(2-D-Pen, 5-D-Pen)-enkephalin, 3H-(-)-ethylketocyclazocine and 3H(D-Ala2-NMePhe-Glyol5)-enkephalin were used as ligands for opioid receptors on neuronal membrane preparations of eyestalk ganglia under consideration of their stereospecific binding properties. In context with saturation binding isotherms, association and dissociation plots, we demonstrate here two opioid receptors; a delta-type receptor with high affinity (Bmax 68.5 fmol/mg protein, Kd = 4.0 nM) and low affinity (Bmax 493 fmol/mg, Kd = 83.6 nM) and a second receptor of kappa-type with Bmax of 3.1 pmol/mg protein and Kd = 68.6 nM.

Dynamics of ocular surface pH.

We studied ocular surface pH in 161 subjects. The mean pH for 133 normal volunteers was 7 . 11, SD 1 . 5. We found that older women had a more alkaline pH than other subjects, that the pH shifted from acid to alkaline during the day, that one hour of eyelid closure caused an acid shift in pH, and that pH recovered to baseline values within 40 minutes after acid drop instillation. We explored the mechanism of pH regulation, and we believe that pH changes could affect contact lens toleration, drug effectiveness, and clinical signs in disease processes.

Isolation and purification of calmodulin from the shrimp, Crangon crangon.

Calmodulin was isolated and purified from shrimp abdominal muscle by heat precipitation, ion exchange and hydrophobic interaction chromatography. The purified calmodulin was homogeneous when evaluated by polyacrylamide gel electrophoresis. A still remaining contaminant was eliminated by high performance liquid chromatography on a phenyl column. The biological and physicochemical properties of shrimp calmodulin such as amino acid composition, molecular weight and the ability to activate calmodulin-deficient bovine heart phosphodiesterase were compared to those of other invertebrate calmodulins.

Immunocytochemical demonstration of the neurosecretory X-organ complex in the eyestalk of the crab Carcinus maenas.

An antiserum was obtained by immunizing rabbits with sinus gland extracts from Carcinus maenas. The antiserum is almost exclusively directed against neurosecretory material in the medulla terminalis X-organ (MTGXO), as demonstrated by the peroxidase--antiperoxidase (PAP) staining method in light and electron microscopic studies. Radioimmunological binding studies indicate the presence of antibodies against the crustacean hyperglycemic hormone (CHH) or the black pigment dispersing hormone (BPDH) in the antiserum. The results suggest that the neurosecretory perikarya of the MTGXO are the sites of production of CHH and/or BPDH.

Tracing of neurosecretory neurons in crayfish optic ganglia by cobalt iontophoresis.

The axonal connections between the medulla terminalis ganglionic X-organ (MTGXO) and the sinus gland are traced by iontophoretic application of cobalt dye to the neurosecretory system in the eyestalks of the crayfish, Orconectes limosus. The MTGXO consists of about 15 large perikarya, forming a distinct subgroup of neurosecretory cells in the medulla terminalis and giving rise to a prominent fibre bundle. Additional axons reaching the sinus gland from the medulla interna, the medulla externa and the optic nerve are less conspicuous.

Strabismus surgery on the nonamblyopic eye.

To date most surgeons operate only on the nonamblyopic eye in patients with strabismus and amblyopia with unpredictable, initial, surgical results. We present a series of 13 patients with amblyopia and strabismus plus anomalous head positions who underwent surgery on their nonamblyopic eye. The good, initial, surgical results in 77% of these patients suggest that operating on the good eye is important to dictate the final surgical results in correcting the deviation and normalizing the head position.

Radioimmunoassay of an invertebrate peptide hormone - the crustacean neurosecretory hyperglycemic hormone.

Antibodies against hyperglycemic hormone (CHH) of Carcinus were raised in rabbits by injection of extract from sinus glands which contain high concentrations of CHH. The antiserum neutralizes the biological activity of CHH and binds 125-J-CHH. A RIA for CHH was established and was used to measure the hormone content of sinus glands.

Immunocytochemical identification of hyperglycemic hormone-producing cells in the eyestalk of Carcinus maenas.

Antiserum raised in rabbits against extracts of sinus glands from Carcinus annd shown by several criteria to contain antibodies directed against the neurosecretory hyperglycemic hormone was used to locate the hormone-producing perikarya in the optic ganglia. By means of the double antibody fluorescence technique, selective staining of the large neurosecretory perikarya of the medulla terminalis ganglionic X-organ (MTGXO) and their axons is obtained. The axon endings of the sinus gland are also stained. None of the other groups of neurosecretory cells in the eyestalk shows fluorescence. Preabsorption of the antiserum with pure hyperglycemic hormone abolishes the fluorescence.

Stromal keratitis with anterior membrane dystrophy.

Anterior stromal nummular infiltrates developed in a 55-year-old woman with recurrent corneal erosion syndrome and associated anterior basement-membrane dystrophy. Gram and Giemsa stains showed no organisms, and cultures revealed no growth at 72 hours. Stromal keratitis can complicate recurrent corneal erosions associated with anterior basement-membrane dystrophy.

Evidence for a crustacean hyperglycemic hormone-like molecule in the nervous system of the stick insect, Carausius morosus.

Histological sections of the brain, suboesophageal ganglion, and the corpora cardiaca/corpora allata complex were examined for the presence of crustacean hyperglycemic hormone-like immunoreactive material. With the use of an antiserum directed against the hyperglycemic hormone of Carcinus maenas, immunofluorescence was found in the median portion of the pars intercerebralis, and the corpora cardiaca. Extracts of corpora cardiaca were examined by radioimmunoassay for competitive binding to the antiserum; one pair of corpora cardiaca contains at least 7 pg crustacean hyperglycemic hormone-like material.

The hyperglycemic neuropeptide of the terrestrial isopod, Porcellio dilatatus. II. Immunocytochemical demonstration in neurosecretory structures of the nervous system.

By use of an rabbit antiserum raised against the crustacean hyperglycemic hormone (CHH) of the shore crab, Carcinus maenas, CHH-producing perikarya were detected in the brain of the isopod, Porcellio dilatatus, by the double-antibody immunofluorescence technique. The course of the secretory axons to the neurohemal organ, the sinus gland, could be traced. The sinus gland also exhibited strong fluorescence. The reaction was very specific, no other structures gave appreciable immunofluorescence. There are two CHH cells in each hemisphere of the brain, located in the medio-rostral part of the protocerebrum. This area appears to correspond to the medulla terminalis of decapods. The cells are aldehyde fuchsin-positive and have been previously described as beta 1 cells. The immunofluorescence could be inhibited by preabsorption of the antiserum with pure Carcinus-CHH.

Biological properties and partial purification of a growth factor from porcine spleen.

3T3-Fibroblast growth is enhanced in a dose-dependent way by a factor isolated from porcine spleen after limited proteolysis. The factor is not dialyzable and is stable for 5 min at 100 degrees C and at pH 2 at room temperature. Trypsin or collagenase treatment does not affect its biological activity. The stimulation of cell growth is independent of serum concentration in the culture medium but is accelerated by insulin supplement. The biochemical properties of this factor indicate a novel growth promotor different from other growth factors.