bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic Diversity of Xanthomonas arboricola pv. fragariae Strains and Comparison with some other X. arboricola Pathovars using Repetitive PCR Genomic Fingerprinting
Authors: Scortichini M. , Rossi M. P.
Published: March 2003

The genetic relationship within 26 Xanthomonas arboricola pv. fragariae strains and between this pathovar and 20 strains of X. arboricola pv. corylina, 22 strains of X. arboricola pv. juglandis and 16 strains of X. arboricola pv. pruni has been assessed by means of repetitive polymerase chain reaction (rep‐PCR) using Enterobacterial Repetitive Intergenic Consensus), BOX (BOXA subunit of the BOX element of Streptococcus pneumoniae) and repetitive extragenic palindromic primer sets. Cluster analysis was performed by means of unweighted paired group method using arithmetic average (UPGMA). Upon rep‐PCR and UPGMA cluster analysis, a relevant genetic diversity was found within the strains. The overall similarity, however, was high (i.e. 80%). The four X. arboricola pathovars showed similar but clearly different genomic patterns and clustered into four different groups, with X. arboricola pv. corylina and X. arboricola pv. juglandis more closely related to X. arboricola pv. fragariae. Representative strains of X. arboricola pv. fragariae and the putative xanthomonads isolated from strawberry leaves showing leaf blight symptoms underwent pathogenicity tests. After artificial inoculation, X. arboricola pv. fragariae induced necrotic spots accompanied, sometimes, by a chlorotic halo. The blackening of the leaf veins and peduncle was, sometimes, also observed. The four putative xanthomonads isolated from diseased strawberry leaves and not inducing symptoms after artificial inoculation, clustered apart from X. arboricola pathovars.
Effects of Sunflower Oil on Tomato Powdery Mildew Caused by Oidium neolycopersici
Authors: Ko W. H. , Wang S. Y. , Hsieh T. F. , Ann P. J.
Published: March 2003

When tomato leaves were sprayed with 0.1% emulsified canola oil, corn oil, grape seed oil, peanut oil, safflower oil, soya bean oil or sunflower oil, the severity of powdery mildew caused by Oidium neolycopersici was greatly reduced. Among these edible oils tested, sunflower oil was the most effective in the control of powdery mildew. When sprayed with 0.5% sunflower oil, powdery mildew on tomato leaves was reduced to a negligible level. Sunflower oil applied to halves of upper leaf surface did not induce resistance against the pathogen in the non‐treated halves. When applied to halves of lower leaf surface, it also failed to reduce the severity of powdery mildew on the upper leaf surface right above the treated area indicating that control of the powdery mildew by sunflower oil did not result from activation of host defence mechanisms. Scanning electron microscopy showed that control of powdery mildew with sunflower oil resulted mainly from the inhibition of conidial germination and suppression of mycelial growth of the pathogen.
Freezing of Stallion Semen with Addition of Glycine Betaine
Authors: Lindeberg H. , Kurtén A. , Koskinen E. , Katila T.
Published: March 2003

The effect of addition of glycine betaine to a lactose‐EDTA freezing medium on the post‐thaw motility of stallion semen was determined. The first three semen‐rich fractions of nine stallions were collected with an open‐end Krakow artificial vagina on consecutive weekdays. Semen was frozen using the Hannover method with freezing media containing glycine betaine in various concentrations from 0 to 5 %. After thawing, sperm motility was analysed both by a light microscope and by a Hamilton‐Thorn Motility Analyser. Total and progressive post‐thaw motilities of semen containing 0.25–3 % glycine betaine did not differ significantly from the total and progressive post‐thaw motilities of semen frozen without glycine betaine. The total and progressive post‐thaw motilities of semen containing 4 or 5 % glycine betaine were significantly lower (P < 0.001) than those of semen without glycine betaine. In conclusion, glycine betaine did not show any beneficial effect on the post‐thaw motility of stallion semen when semen was frozen using the Hannover method.
Effects of Plant‐Induced Hypervitaminosis D on Cutaneous Structure, Cell Differentiation and Cell Proliferation in Cattle
Authors: Gimeno E. J. , Costa E. F. , Gomar M. S. , Massone A. R. , Portiansky E. L.
Published: March 2003

Solanum glaucophyllum (Sg) (synonym S. malacoxylon) is a plant toxic to cattle due to its high levels of 1,25‐dihydroxyvitamin D3 as glycoside derivatives. Sg causes a disease characterized by wasting and calcification of soft tissues. The effects of vitamin D are not only important in calcium homeostasis, but also in immune regulation, cell growth and cell differentiation. Skin samples in Sg‐intoxicated and control heifers were studied histologically. Cellular differentiation and proliferation were analysed by immunohistochemical expression of cytokeratins, involucrin and proliferating cell nuclear antigen (PCNA). The results were obtained by image processing and analysis and were statistically evaluated. Sg‐intoxicated cattle showed atrophy of epidermis and severe involution of hair follicles and of sebaceous and sweat glands. As judged by PCNA expression, cellular proliferation was reduced, even though the reduction was not statistically significant. The analysed markers of differentiation, e.g. involucrin and cytokeratins 10 and 11, changed in relation to Sg‐poisoning. The possible pathogenesis of the skin lesions is discussed.
Prevention of Scours in Neonatal Kids after Oral Administration of an Organic Acid Solution
Authors: KRITAS S. K.
Published: March 2003

The purpose of this study was to evaluate under field conditions, the efficacy of mild organic acid solutions in the prevention of neonatal kid diarrhoea. At a goat farm, two experimental groups of approximately 120 kids each were formed. The kids of the first group were not submitted to any treatment and served as negative controls, whereas the kids of the second group received a solution of organic acids (Euroacid 50‐L; Eurotec, Waterloo, Belgium) which was administered orally on the first and second day of life. Groups were compared with regard to the incidence of diarrhoea, its duration, and the mortality of the kids. The results showed that, in comparison with the control group, the morbidity, the mortality and the case fatality of the treated kids was significantly reduced (P < 0.05). Furthermore, the duration of diarrhoea per sick kid was markedly reduced in the acidifier‐treated group in comparison with the control animals (P < 0.05). Cultures of the diarrhoeic faeces from kids indicated that enterotoxigenic strains of Escherichia coli, and more specifically strains that were positive for K88 and K99 antigens, were present in the particular farm. It was concluded that the administration of organic acids can be a helpful means in controlling scours in neonatal kids.
Haemangiosarcoma with a Metastasis of a Malignant Mixed Mammary Gland Tumour in a Dog
Authors: HILBE M. , HAUSER B. , ZLINSZKY K. , EHRENSPERGER F.
Published: March 2003

An 11‐year‐old dog with a history of weight loss, anorexia, anaemia, epistaxis and a nodule in the mammary gland showed multifocal lung masses during radiography. At necropsy as well as in the histological examination two different tumours were identified in several organs including the right heart auricle. By immunohistochemistry, using a double‐staining DAKO® kit (DAKO Diagnostics, Zug, Switzerland), it was possible to identify the two components as haemangiosarcoma and metastasizing malignant mixed mammary gland tumour.
PrP Genotyping of Austrian Sheep Breeds
Authors: SIPOS W. , KRAUS M. , SCHMOLL F. , ACHMANN R. , BAUMGARTNER W.
Published: March 2003

Scrapie, an ovine and caprine transmissible spongiforme encephalopathy, is widely spread among sheep populations in many European countries. As it is known that susceptibility to scrapie is determined genetically, breeding programmes aiming at providing scrapie‐resistant flocks have been established. Selection is based on the prion protein (PrP) genotype, which is used to classify animals into risk groups of susceptibility (R1–R5) according to the amino acids encoded by codons at positions 136, 154 and 171, respectively. At position 136 (136V→136A) alanine and at position 154 (154H→154R) as well as 171 (171Q→171R) arginine are the favoured amino acids. Whereas PrP genotyping data are available for many of the European sheep breeds, comparable data for local Austrian sheep breeds are missing. The most known among these are Tyrolean mountain sheep, forest sheep, Tyrolean stone sheep and Carynthian sheep. The genotypes of 112 sheep from these four local breeds were determined. In terms of PrP genetics, Austrian breeds belong to the group of non‐valine‐breeds, with the exception of the Carynthian sheep, that exhibited a frequency of 136V of 4.2%. The most frequent allele was ARQ with 64.6–71.2% (depending on the breed), followed by ARR (14.8–25.8%). In contrast to the above‐mentioned findings, scrapie has never been diagnosed in any of the Austrian sheep breeds. Native Austrian sheep breeds exhibit a very robust constitution, a pronounced adaptation to harsh climates and good reproduction parameters as well as a marked mother‐instinct. Therefore, these breeds are often used in cross‐breeding programmes. Beside the above‐mentioned characteristics, our results indicate that the investigated breeds may be effectively used in crossing‐out breeding programmes for eliminating valine at position 136 of PrP.
Rumen‐Protected Choline Administration to Transition Cows: Effects on Milk Production and Vitamin E Status
Authors: Pinotti L. , Baldi A. , Politis I. , Rebucci R. , Sangalli L. , Dell'Orto V.
Published: March 2003

We investigated the influence of rumen‐protected choline (RPC) supplementation on milk production, lipid metabolism and vitamin E status in dairy cows receiving a silage‐based diet. Twenty‐six Italian Holstein multiparous cows were assigned by weight and average production in the previous lactation, to one of two groups: control (no RPC supplementation) and RPC (supplemented with 20 g/day rumen‐protected choline chloride). Treatment began 14 days before expected calving and continued for 30 days after parturition. Choline administration significantly increased milk production during the first month of lactation and also the concentration (and total secretion) of choline in milk, but did not affect fat or protein concentrations in milk, or plasma levels of glucose, β‐hydroxybutyrate, cholesterol and non‐esterified fatty acids (NEFA). However, around parturition, NEFA concentrations in plasma were lower in treated animals than in controls, suggesting improved lipid metabolism as a result of choline supplementation. Choline supplementation also increased α‐tocopherol plasma concentrations, suggesting a novel aspect in dairy cows.
Evaluation of Selective Media for Bifidobacteria in Poultry and Rabbit Caecal Samples
Authors: Rada V. , Sirotek K. , Petr J.
Published: March 2003

Five media were evaluated to determine their selectivity for Bifidobacterium sp. in hen and rabbit caecal samples. The colonies arising on the plates inoculated with the caecal samples were Gram stained and screened for the presence of fructose‐6‐phosphate phosphoketolase activity. Rogosa agar modified by the addition of cysteine‐hydrochloride (0.05 % w/v), Beeren’s agar (with 5 ml/l of propionic acid as a selective agent), BS 2 agar (containing per one litre sodium propionate 15 g, lithium chloride 3 g, paromomycin sulphate 50 mg, neomycin sulphate 200 mg), and Wilkins‐Chalgren agar (MW) modified by the addition of acetic acid (1 ml/l) and mupirocin (100 mg/l) were selective for Bifidobacterium sp. from rabbit caecal samples. In contrast, only MW medium was suitable for the isolation and enumeration of bifidobacteria in hen caecal samples. In conclusion, the results suggest that MW agar showed the greatest selectivity. A further advantage of this medium is its ease of preparation. Therefore this agar could contribute to the study of the effects of the ingestion both probiotics and prebiotics. Finally, it could be noted that the bifidobacteria selective media should be chosen in respect of the animal species origin of the sample tested.
Institute of Animal Experimentation, Sapporo Medical University, Minami 1 Nishi 17, Sapporo 060, Japan
Authors: Isogai H. , Kosako Y. , Benno Y. , Isogai E.
Published: March 2003

Asaccharolytic pigmented Porphyromonas species, including P. endodontalis, P. gingivalis, P. circumdentaria and unclassified species, were isolated from the plaque of adult dogs, but not from any oral sites of puppies and adolescent dogs. With age‐dependency, the proportion of Porphyromonas species in the flora of plaque increased. Isolation of the genus Porphyromonas was clearly associated with the progress of periodontol disease. We suggested that Porphyromonas is the exogenous organism and obligate pathogen for canine periodontal diseases.
Restriction endonuclease analysis of BHV‐1 and BHV‐5 strains isolated in argentina
Authors: Pidone C. L. , Galosi C. M. , Echeverria M. G. , Nosetto E. O. , Etcheverrigaray M. E.
Published: March 2003

The genomes of 10 bovine herpesvirus 1 and 5 strains isolated in Argentina from 1989 to 1994, recovered from animals showing different clinical signs, and two reference strains (Los Angeles and A663) were compared by restriction endonuclease analysis. Four restriction enzymes, HindIII, BamHI, EcoRI and PstI, were used and analysis of the restriction patterns used to assign the isolate to either the BHV‐1.1, BHV‐1.2 or BHV‐5 genotype. There was a correlationship between restriction pattern and clinical signs in six out of ten Argentinian isolates.
Cow–Calf Herds in Eastern Germany: Status Quo of Some Parasite Species and a Comparison of Chemoprophylaxis and Pasture Management in the Control of Gastrointestinal Nematodes
Authors: Wacker K. , Roffeis M. , Conraths F. J.
Published: March 2003

Infections with gastrointestinal parasites (Eimeria spp., Cryptosporidium spp., Buxtonella sulcata, Fasciola hepatica, Moniezia spp. and trichostrongyles) and lungworms were monitored in five cow–calf herds in the north German lowlands. Estimated prevalences of infections with Eimeria spp. (predominantly Eimeria bovis) ranged between approximately 2 and 48 %. The highest prevalences were found during late summer and autumn. On one farm Cryptosporidium spp. were detected in July and August (prevalence: 8.5 ± 2.7 % and 6.7 < 2.0 %). The latter finding coincided with diarrhoea in many calves. Buxtonella sulcata was found during the entire study period in highly variable estimated prevalences ranging between zero and 73 %, but without any obvious association with clinical disease. Fasciola hepatica was detected on four out of five farms at estimated prevalences of approximately 1–20 %. Lungworm infections played a minor role in at least three of five study herds. The estimated prevalence of trichostrongyle infections rose from August until November whereas the intensity of infection did not change significantly. No difference in the intensity of infection could be detected between one farm on which infections with gastrointestinal nematodes were controlled only by moving the animals to an uninfected pasture in July, and three other herds on which strategic anthelmintic control was in place.
Immunohistochemical Detection of Antigen in Lamb Tissues Naturally Infected with Sheeppox Virus
Authors: Gulbahar M. Y. , Cabalar M. , Gul Y. , Icen H.
Published: March 2003

The present study describes the detection of sheeppox virus antigen in various lamb tissues, using an immunohistochemical technique, in sheeppox cases which occurred naturally. Sheeppox viral antigen was detected in the cytoplasm of sheeppox cells and degenerated epithelial cells of the skin, lungs and digestive tract involving typical sheeppox lesions. Nuclear staining was also observed in some typically deformed nuclei of sheeppox cells. The immunostaining of sheeppox virus showed a correlation with the presence of sheeppox cells and degenerated epithelial cells resembling them. Additionally, in order to confirm the presence of sheeppox virus in the skin and lung samples, direct electron microscopy was performed and sheeppox virus was only demonstrated in two skin samples.
Production and Characterization of a Panel of Monoclonal Antibodies for the Identification of Antigens of Babesia bovis
Authors: Figueiredo J. F. , Martins M. S. , Ribeiro M. F. B. , Passos L. M. F.
Published: March 2003

A panel of monoclonal antibodies was produced and characterized by an indirect fluorescent antibody test (IFAT), an enzyme‐linked immunosorbent assay (ELISA) and Western blotting with the aim of identifying antigens of Babesia bovis. After fusion, the resultant hybrids were selected by the IFAT, cloned, maintained in culture in vitro, and cryopreserved in liquid nitrogen. Ten clones producing monoclonal antibodies were found to react against the entire merozoites, three reacted on the surface of the merozoites, and one clone reacted against the polar region of the merozoites. All monoclonal antibodies reacted in ELISA, with the optical density varying from 0.368 to 0.502 (cut off = 0.022). The bands recognized by the monoclonal antibodies in Western blotting had molecular weights ranging from 162 to 19 kDa. Four clones recognized a single band of 73 kDa, and another four did not react in Western blotting.
Prevalence of Antibodies to Leptospira Serovars in Sheep and Goats in Alto Adige–South Tyrol
Authors: Ciceroni L. , Lombardo D. , Pinto A. , Ciarrocchi S. , Simeoni J.
Published: March 2003

Serum samples from 313 sheep and 95 goats were collected during November 1993 in 26 localities in Alto Adige–SouthTyrol and tested by microscopic agglutination test for antibodies to 28 serovars of the genus Leptospira. At the time of blood collection all the animals appeared healthy with no clinical sign suggestive of leptospirosis. The observed seroprevalence in sheep was 6.1 %, whereas the seropositivity rate for goat serum samples was 2.1 %. The highest serological prevalence in sheep was recorded for serovar castellonis, followed by poi, sejroe, hardjo subtype hardjobovis, copenhageni, and cynopteri. Titres to poi were the only ones found in goats. These findings, which are proof of Leptospira infection in Alto Adige–South Tyrol, indicate that foci of several serovars exist in this region.
The Sensitivities of Various Erythrocytes in a Haemagglutination Assay for the Detection of Psittacine Beak and Feather Disease Virus
Authors: Sanada N. , Sanada Y.
Published: March 2003

The erythrocytes of various species were tested in psittacine beak and feather disease (PBFD) virus haemagglutination (HA) and haemagglutination inhibition assays to determine which are suitable for use in these assays. HA activity was observed for erythrocytes of the salmon‐crested cockatoo, the sulphur‐crested cockatoo, the umbrella cockatoo, the goffin's cockatoo and the cockatiel, with differences amongst individuals within species, but not for erythrocytes of humans, the pig, the guinea pig, the chicken, the goose, the rose‐ringed parakeet or the budgerigar. Anti‐PBFD virus rabbit sera inhibited the virus‐induced agglutination of erythrocytes, confirming the specificity of HA activity. This suggests that selection of suitable psittacine species as well as suitable individuals within a species is necessary when obtaining erythrocytes for the PBFD virus HA assay.
A Polymerase Chain Reaction for Detection of Equine Herpesvirus‐1 in Routine Diagnostic Submissions of Tissues from Aborted Foetuses
Authors: Galosi C. M. , Roza M. V. Vila , Oliva G. A. , Pecoraro M. R. , Echeverría M. G. , Corva S. , Etcheverrigaray M. E.
Published: March 2003

Equine herpesvirus 1 (EHV‐1) is the causative agent of abortion, perinatal foal mortality, neurological and acute respiratory diseases in horses. Conventional laboratory diagnosis involving viral isolation from aborted foetuses is laborious and lengthy and requires processing of samples within 24 h of collection, which is problematic for samples that come from long distances. The aim of this study was to develop a polymerase chain reaction (PCR) assay useful in Argentina to detect DNA sequences of EHV‐1 in different tissues from aborted equine foetuses with variable quality of preservation and without the use of conventional DNA fenolic extraction. Several DNA extraction protocols and primers were evaluated. The amplification method was standardized and its specificity was analysed using 38 foetal samples of variable quality of preservation. Of the 38 different foetal tissues, nine livers, six spleens and two lungs in good preservation and eight livers, one spleen and four lungs in a poor state of preservation were positive for PCR. EHV‐1 was recovered only from the nine livers, five spleens and two lungs in good preservation. No virus was isolated from the samples that were poorly preserved. Viral isolation was confirmed by cytopathic effect and indirect immunofluorescence. The specificity of the PCR results was confirmed by the restriction endonuclease digestion of PCR products and hybridization.
Studies on Streptococcus agalactiae Isolated from Bovine Mastitis in Indonesia
Authors: ESTUNINGSIH S. , SOEDARMANTO I. , FINK K. , LÄMMLER C. , WIBAWAN I. W. T.
Published: March 2003

All 83 bacterial strains isolated from seven farms in three areas of the island of Java in Indonesia investigated in the present study could be identified as Streptococcus agalactiae. Identification was performed by cultural, biochemical and serological properties and by polymerase chain reaction amplification of species‐specific parts of the gene encoding the 16S rRNA, the 16S–23S rDNA intergenic spacer region and the CAMP factor (cfb) gene. All isolates were unpigmented, almost all of the isolates had the serotype pattern II/X. Despite these similarities a macrorestriction analysis of the chromosomal DNA of the bacteria revealed no significant homologies of the DNA‐fingerprints of the S. agalactiae from the various areas. This last finding might possibly indicate that a single ancestral unpigmented serotype II/X S. agalactiae clone was responsible for the mastitis situation on Java and had evolved separately in the various farms and regions.
Bacterial Colonization and Invasion in Pigs Experimentally Exposed to Streptococcus suis Serotype 2 in Aerosol
Authors: MADSEN L. W. , BAK H. , NIELSEN B. , JENSEN H. E. , AALBÆK B. , RIISING H. J.
Published: March 2003

A recently developed porcine model for aerogenous infection with Streptococcus suis serotype 2 was applied in a study of the phases of bacterial colonization and initial invasion. Eighteen pigs were exposed to aerosolized S. suis serotype 2 after pre‐exposure to mild acetic acid in aerosol. The animals were killed consecutively within the first six days after challenge. After death, all animals were necropsied and examined by bacteriology, histopathology, and immunohistochemistry.

Systemic infection was established in four out of 18 animals exposed to S. suis serotype 2. All systemically infected animals developed clinical signs and lesions typical of the infection. In four additional animals, subclinical infection was demonstrated by re‐isolation of S. suis from the palatine tonsil. However, in all 18 challenged animals, immunohistochemistry demonstrated S. suis serotype 2 antigen in the palatine and/or nasopharyngeal tonsils. In all four systemically infected animals, S. suis serotype 2 antigen was also found in the mandibular lymph node. These observations point towards the tonsils as possible portals of entry for S. suis serotype 2 with subsequent lymphogenous spread. Thus, the present findings parallel the proposed pathogenesis for S. suis serotype 1 infection in pigs.
Specific Detection of Chikungunya Virus Using a RT‐PCR/Nested PCR Combination
Authors: PFEFFER M. , LINSSEN B. , PARKER M. D. , KINNEY R. M.
Published: March 2003

Chikungunya (CHIK) virus is enzootic in many countries in Asia and throughout tropical Africa. In Asia the virus is transmitted from primates to humans almost exclusively by Aedes aegypti, while various aedine mosquito species are responsible for human infections in Africa. The clinical picture is characterized by a sudden onset of fever, rash and severe pain in the joints which may persist in a small proportion of cases. Although not listed as a haemorrhagic fever virus, illness caused by CHIK virus can be confused with diseases such as dengue or yellow fever, based on the similarity of the symptoms. Thus, laboratory confirmation of suspected cases is required to launch control measures during an epidemic. CHIK virus diagnosis based on virus isolation is very sensitive, yet requires at least a week in conjunction with virus identification using monovalent sera. We developed a reverse transcription–polymerase chain reaction (RT‐PCR) assay which amplifies a 427‐bp fragment of the E2 gene. Specificity was confirmed by testing representative strains of all known alphavirus species. To verify further the viral origin of the amplicon and to enhance sensitivity, a nested PCR was performed subsequently. This RT‐PCR/nested PCR combination was able to amplify a CHIK virus‐specific 172‐bp amplicon from a sample containing as few as 10 genome equivalents. This assay was successfully applied to four CHIK virus isolates from Asia and Africa as well as to a vaccine strain developed by USAMRIID. Our method can be completed in less than two working days and may serve as a sensitive alternative in CHIK virus diagnosis.
An Unusual Outbreak of Malignant Catarrhal Fever in a Beef Herd in Israel
Authors: Brenner J. , Perl S. , Lahav D. , Garazi S. , Oved Z. , Shlosberg A. , David D.
Published: March 2003

Malignant catarrhal fever (MCF, corrizza contagiosa) is an invariably fatal communicable disease in cattle, whose causative agent is the ovine herpes virus‐2, or the alcelaphine herpes virus‐1. In one feed‐lot family farm, 34 calves out of 100 became ill at the rate of one to four calves per week, and all of them subsequently died over a period of 4 months. Most of the initial cases were manifested clinically as the head and eye form, but most of the entire clinical spectrum of forms (the respiratory, intestinal and nervous forms) characteristic for MCF were observed as this epidemic progressed. Very few calves died without showing any specific signs of MCF. Pathological examinations revealed characteristic obliterative arteriovasculitis in the brain of calves with nervous signs, typical of MCF. Polymerase chain reaction (PCR) testing revealed 100% homology between the 238 bp hemi‐nested PCR fragment and the ovine herpes virus‐2 sequences.

Based on the clinical signs, epidemiological data, pathological, and histopathological findings, and the PCR results, it was concluded that MCF occurred on the farm. The fact that sheep and goats were housed in close proximity on the same farm reinforced this diagnosis.
Optimization of a Culture Technique for the Isolation of Listeria monocytogenes from Faecal Samples
Authors: Erdogan H. M. , Cripps P. J. , Morgan K. L.
Published: March 2003

A culture technique employing cold enrichment at 4°C followed by selective enrichment and plating at higher temperatures (30°C) was used to isolate Listeria monocytogenes from faecal samples. The samples were held at 4°C for 15 weeks and cultured weekly to assess the sensitivity of the culture after cold storage for different lengths of time. No media, Listeria selective enrichment broth (LSEB), nutrient broth (NB) and saline were used as cold storage medium. Cold storage increased the frequency of Listeria positive samples. The sensitivity of the culture for Listeria spp. and L. monocytogenes was 72 and 94%, and 56 and 61% after third and seventh week of cold storage, respectively. When the results of third and seventh week of cold storage were combined, the sensitivity was 100% for Listeria spp. and 94% for L. monocytogenes. LSEB and NB as storage medium increased Listeria positive samples after the first week of cold storage but did not maintain the increase thereafter while saline had an adverse effect on the growth of the bacteria. However, samples held in no media in a pilot study involving monthly sampling of a herd revealed better results. Detection limit of the culture media was also investigated. The lowest concentration detected by culture media was 3.17 organisms/ml. This was seven organisms/g for known Listeria positive sample. The faecal samples spiked with 10‐fold dilutions of L. monocytogenes and held at 4°C revealed that the sample spiked with 3.17 × 10−1 cfu/ml organisms resulted in growth after the second week of cold storage. The results suggest that the culture technique employing cold enrichment followed by selective enrichment and plating is more sensitive, the storage of faecal samples in no media when compared with the samples in storage medium, LSEB, NB and saline, during cold enrichment is a better application and culture of faeces, immediately after collection, at third and seventh week of cold enrichment produce more satisfactory results.
An In‐Situ Study on the Influence of Ascidian Suspension Feeding on the Subtidal Nepheloid Layer in the Northern Adriatic Sea
Authors: Hartl Mark G. J. , Ott Jörg A.
Published: March 2003

Abstract. In order to evaluate the effect of epibenthic suspension‐feeding organisms on nepheloid‐layer processes, a novel flow‐through ‘Benthic Ecosystem Tunnel’ was used on sublittoral marine sediment substrate. Our test organism was the ascidian Microcosmus sulcatus. Particle and chlorophyll a reduction were recorded. The correlation of seasonal fluctuations of chlorophyll a concentrations with chlorophyll a reduction by M. sulcatus indicates the potential for Microcosmusto control local phytoplankton input to the subtidal nepheloid layer. However, the relatively low density of Microcosmus at the study site, the nature of the sediment surface and the susceptibility of ascidians to high turbidity (i.e., high sedimentation rates of total particular matter (TPM) causing frequent periods of siphon closure and inactivity) makes it unlikely that M. sulcatusis an important element controlling the composition of the nepheloid layer on a large scale.
The Effects of Hermit Crabs  on Hydractiniid Hydroids The effects of hermit crabs on hydractiniid hydroids
Authors: Winkle David H. Van , Longnecker Krista , Blackstone Neil W.
Published: March 2003

Abstract. Hermit crab shells are often encrusted with diverse communities of epibionts. To explore the effects of hermit crabs on the interactions of these encrusting species, two species of hydractiniid hydroids, Hydractinia symbiolongicarpus and Podocoryna carnea, were examined in the presence and absence of Pagurus longicarpus hermit crabs. Colonies growing on Littorina littorea snail shells with hermit crab hosts were compared to those growing on shells without crabs. These experiments suggest that hermit crabs variably affect colony polyp number, colony morphology and the outcome of interspecific competition, according to the size and number of crabs; other factors include water temperature, food availability and hydroid clonal genotype. In order to allow image analysis of perturbations of colony morphology, hydroids were grown on glass surfaces both exposed and unexposed to hermit crabs. Relative to colonies growing on unexposed surfaces, colonies growing on exposed surfaces exhibited larger within‐colony patches, thus decreasing the total encrusted surface area and polyp number of colonies. Further, on glass surfaces, hermit crabs accelerated the time to the resolution of interspecific competition (i.e., the overgrowth of one colony by the other). While a number of factors may contribute to the relative abundance of these encrusting species, under most circumstances P. longicarpus have large effects on hydractiniid hydroids. Mechanisms underlying these effects likely include hydrodynamics and mechanical disturbance.
Deep‐Sea Diversity:  Overlooked Messages from  Shallow‐Water Sediments
Authors: Grant Alastair
Published: March 2003

Abstract. Much of the discussion of high deep‐sea diversity has assumed that asymmetric inter‐specific competition will rapidly lead to the elimination of many species unless restrained by extensive differentiation of niches, or the action of predation and/or environmental disturbance. This is true for some habitats, including rocky shores. However, experimental studies indicate that marine soft sediment communities do not function like this. In shallow‐water sediments, competition is usually symmetric and relatively weak. Asymmetric competition that leads to the elimination of one species by another on time scales shorter than one generation is rare, apart from interactions that involve large‐scale modification or disturbance of the sediment. Competition is therefore relatively unimportant as a process structuring communities and the impact of predation is usually to reduce rather than enhance diversity. These results have been largely ignored by the literature on deep‐sea diversity. If deep‐sea communities function in similar ways, coexistence of many species within small areas on short time scales does not need further explanation. We do, however, need to explain why abundances of all species remain bounded on large spatial scales and time scales of several generations. The model of diversity maintenance proposed by
Huston (1979) and applied to the deep sea by
Rex (1983) achieves this by implicit intra‐specific, density‐dependent processes that increase the population growth rate of species that have become locally rare. This would give robust maintenance of diversity, but there is no evidence for density dependence of this type in the deep sea, and no plausible mechanisms by which it could occur. Alternative models require either spatial heterogeneity on a scale much larger than that envisaged by the grain‐matching hypothesis or the placing of a cap on the abundance of common species, perhaps by frequency‐dependent predation. Arbitrating between these possibilities will require assessments of the population dynamics and spatial distribution of individual species on spatial and temporal scales much greater than those usually considered in the deep sea.
Spatial Overlap and Coexistence in a Mussel‐Associated Polychaete Assemblage on a South American Rocky Shore
Authors: Tokeshi Mutsunori , Romero Leonardo
Published: March 2003

Abstract. Mussel beds in the intertidal of subtropical South America are known to harbour a large number of invertebrates, particularly polychaetes. This paper presents a quantitative analysis of the patterns of spatial overlap and coexistence in an assemblage of polychaetes associated with mussel beds in a rocky intertidal habitat on the Pacific coast of Peru. Mussel beds are made up of two zones in the intertidal: the lower, more extensive zone formed by Semimytilus algosus and the upper zone formed by Perumytilus purpuratus. Null models were employed to assess the magnitude of spatial overlap between pairs of taxa encompassing a total of seven taxonomic groups. Two taxa belonging to the same functional group tended to be spatially segregated in the Semimytilus bed: Pseudonereis vs. Halosydna (large free‐ranging predators) and Scoloplos vs. Mediomastus (deposit feeders), with further segregating tendencies between Pseudonereis and Mediomastus, and between Halosydna and Scoloplos. Small‐sized, free‐ranging ‘Typosyllis’ and the bottom‐dwelling scavenging predator Lumbrineris were also spatially associated. Thus, this polychaete assemblage appears to be loosely organised around groupings of taxa which are somewhat different in ecological traits and form mosaic patterns of distribution with reduced overlap within the Semimytilus bed. These relationships were weak to non‐existent in the upper, drier Perumytilus habitat where polychaete density was low.
Enzymatic Approach to Fungal Association with Arthropod Guts: A Case Study for the Crustacean Host,Nihonotrypaea harmandi, and Its Foregut Fungus,Enteromyces callianassae
Authors: Kimura Hiroshi , Harada Kazuyuki , Hara Kenji , Tamaki Akio
Published: March 2003

Abstract.In the callianassid shrimpNihonotrypaea harmandi(Bouvier 1901), the trichomycete fungusEnteromyces callianassaeLichtwardt 1961 occurs exclusively on the foregut lining. The enzymes from both the shrimp and the fungus apparently hydrolyze certain nitrogen and carbon compounds in detritus. The activities of various proteases and carbohydrases contained in the foregut juice were compared between fungus‐infected shrimps collected from a sandflat in Kyushu, Japan (51 % infection rate) and uninfected shrimps from a nearby sandflat. The concentration of enzymatically hydrolyzable amino acids (EHAA) in sediment liberated by the foregut juice with fungi was slightly lower than that by 0.1 mg·ml‐1proteinase‐K and significantly higher than that by the foregut juice without fungi. In both foregut juices, substantial enzyme activities were recorded for proteinase(s), peptidases, amylase, cellulase and β‐1,3‐glucanase, and minimal ones for maltase and cellobiase. Of the commercial substrates examined, only in the case of the mixture of 16 kinds of dipeptides was a significantly higher enzyme activity in the foregut juice with fungi observed. In the process of EHAA liberation from sediment, peptidases secreted by the fungus most probably act as a supplement to the endogenous secretion by the shrimp. This may explain the higher shrimp growth rate recorded for the population with fungi. However, the significantly higher sediment EHAA concentration of the sandflat inhabited by this population versus the population without fungi could be another crucial factor explaining the site difference in shrimp growth rate.
Molecular differentiation of dermatophyte fungi
Authors: Harmsen D. , Schwinn A. , Bröcker E.‐B. , Frosch M.
Published: March 2003

Our previous studies of the ribosomal DNA variation in dermatophytes have shown that these fungi are monophyletic in origin. However, this approach did not allow us to differentiate all the species defined by classical means. Therefore, we studied the internal transcribed spacer 1 (ITS 1) region of 17 species of the fungal order Onygenales, comprising the pathogenic keratinophilic fungi. Interspecific nucleotide composition and sequence length variation of the ITS 1 region was high, mean identities were as low as 40% and sequence lengths varied from 169 to 293 basepairs. Each established dermatophyte species could be identified. In contrast, the flanking sequences at the 3′ end of 18S and the 5.8S rDNA were conserved. Although the value of the ITS 1 region as a phylogenetic tool may be limited because of its high variability, it provides the information necessary to design species‐specific probes, or polymerase chain reaction restriction fragment polymorphism systems useful for taxonomic or rapid diagnostic tests.
Clinicians’ reaction to positive urine culture for Candida organisms
Authors: Ayeni  , Riederer  , Wilson  , Khatib
Published: March 2003

Clinicians’ reaction to isolating Candida organisms in urine culture (≥104 CFU ml−1) was assessed in a retrospective review of 133 consecutive in‐patients (≥15 years‐of‐age) over a 5 month period. The average age was 68.8 years and male/female ratio was 0.36 (35/98). Most (78.2%) patients had an indwelling catheter, and many (35.3%) were in the intensive care unit (ICU). In response to culture‐result, clinicians initiated antifungal therapy in 80 instances (60.2%). Treatment was often based on a single culture without documenting the infection (n=53/80, 66.3%) in the absence of risk for invasive disease. Removing the indwelling‐catheter was never attempted and antibiotics were rarely discontinued or modified (1.3%). Fluconazole was most frequently utilized (n=42, 52.5%), followed by amphotericin‐B bladder‐irrigation (n=26, 32.5%), and combined fluconazole/amphotericin‐B bladder‐irrigation (n=12, 15%). Therapy was more frequently initiated in ICU‐cases (76.6 versus 55.6%; P=0.023) and less often in non‐catheterized individuals (40.7 versus 69%; P=0.012) and patients with 104 CFU ml−1 (25.9 versus 72.7%; P<0.0001). These findings show that clinicians nowadays do not follow current guidelines for the management of candiduria. Efforts to increase clinicians’ awareness of these guidelines, which are intended to confirm the diagnosis and stratify treatment according to patient risk factors, appear to be necessary.
Case Report. Atypical cutaneous pseudallescheriosis refractory to antifungal agents
Authors: Ginter Gabriele , Petutschnig B. , Pierer G. , Soyer H. P. , Reischle S. , Kern T. , De Hoog S.
Published: March 2003

We report on a 65‐year‐old male heart transplant recipient who was otherwise in good condition. The patient was immunocompromised secondarily due to cyclosporin, prednisolone and azathioprine when widespread pustular skin lesions with erythematous margins subsequently developed on his left forearm. There was no history of trauma or septic temperature. Bacterial cultures were sterile and the results of native and cultural investigation studies were negative. A biopsy specimen of the lesion demonstrated hyalohyphomycosis with numerous septate hyphae within granulomas throughout the dermis. Subcutaneous tissues were not involved. Culture plates inoculated with pus and skin from the punch biopsy showed growth of a mould yielding Pseudallescheria boydii. Sensitivity testing was performed with miconazole, ketoconazole and itraconazole showing the best in vitro activity against P. boydii. In spite of treatment with itraconazole, the erythema and pustules continued to spread and therapy was changed to intravenous miconazole. Due to ongoing progression after 3 months of antifungal therapy surgical debridement was required. After 2 years of follow up, he had no recurrence.
Epidemiology of dermatomycoses of humans in Central Poland. Part III. Tinea pedis
Authors: Lupa S. , Seneczko F. , Jeske J. , Głowacka Anna , Ochęcka‐Szymańska Aldona
Published: March 2003

The total number of dermatophytoses (7393) included 2025 (27.4%) tinea pedis cases. Etiological factors in descending order by contribution were: Trichophyton rubrum (41.7%), Trichophyton mentagrophytes var. granulosum (30.9%), T. mentagrophytes var. interdigitale (10.0%), Epidermophyton floccosum (7.4%), T. mentagrophytes var. quinckeanum (6.9%), Trichophyton tonsurans (2.3%), Trichophyton spec.(0.4%), Trichophyton terrestre (0.2%), Trichophyton violaceum (0.1%). In the years 1987–93 the incidence of tinea pedis substantially increased over 1994–96. Today tinea pedis is second by incidence among all clinical forms of dermatophyte infections of skin and skin appendages in the Łódź region.
Intermittent pulse therapy with itraconazole for onychomycosis. Experience in Hong Kong Chinese
Authors: Tang  , Chong  , Leung  , Ho  , Wong
Published: March 2003

Forty‐four Hong Kong Chinese adults comprising 10 cases of fingernail and 42 cases of toenail onychomycosis were recruited for a 3‐weekly itraconazole pulse therapy. Each pulse consisted of seven consecutive daily 400 mg doses with a 3‐week interval between treatments. All patients in the fingernail group and 37 in the toenail group completed the study. The clinical cure, clinical response and mycological cure rates at week 36 were 70, 90 and 90% for fingernail and 35, 81 and 68% for toenail groups, respectively. Side‐effects including biochemical abnormalities were minimal, and returned to normal upon cessation of treatment. We conclude that itraconazole pulse therapy is very effective, safe and well‐tolerated for fingernail and toenail onychomycosis.
Karyotyping of Candida albicans and Candida glabrata from patients with Candida sepsis
Authors: Klempp‐Selb  , Rimek  , Kappe
Published: March 2003

The aim of this study was to determine the relatedness of Candida strains from patients suffering from Candida septicaemia by typing of Candida isolates from blood cultures and different body sites by pulsed field gel electrophoresis (PFGE using a contour‐clamped homogenous electric field, CHEF). We studied 17 isolates of Candida albicans and 10 isolates of Candida glabrata from six patients. Four patients suffered from a C. albicans septicaemia, one patient from a C. glabrata septicaemia, and one patient had a mixed septicaemia with C. albicans and C. glabrata. Eight isolates from blood cultures were compared with 19 isolates of other sites (stool six, urine four, genital swab four, tip of central venous catheter three, tracheal secretion one, sputum one). PFGE typing resulted in 10 different patterns, four with C. albicans and six with C. glabrata. Five of the six patients had strains of identical PFGE patterns in the blood and at other sites. Seven isolates of a 58‐year‐old female with a C. glabrata septicaemia fell into five different PFGE patterns. However, they showed minor differences only, which may be due to chromosomal rearrangements within a single strain. Thus it appears, that the colonizing Candida strains were identical to the circulating strains in the bloodstream in at least five of six patients.
Disc diffusion test in the identification of Candida species
Authors: Menon Thangam , Ponnuvel
Published: March 2003

A total of 448 clinical isolates and seven reference strains of Candida species were tested for their susceptibility to a group of dyes and chemicals. All strains were also identified by conventional methods. The chemicals/dyes used in the disc diffusion tests were brilliant green, janus green, cycloheximide, fast green, rhodamine 6 G, ethidium bromide and 2, 3, 5 triphenyl tetrazolium chloride. Sensitivity to the chemicals was coded by a seven‐figure code. Resistogram patterns obtained were found to be specific for each species. The disc diffusion test was found to be inexpensive, quick and gave reliable results within 18 h.
The frequency of Candida parapsilosis in onychomycosis. An epidemiological survey in Israel
Authors: Segal R. , Kimchi A. , Kritzman A. , Inbar R. , Segal And Z.
Published: March 2003

Candida albicans is regarded as the major pathogen in yeast‐induced onychomycosis. Based on our impression of an increasing prevalence of Candida parapsilosis in this disease, we examined the data of two mycology laboratories in the same geographic location, from 1994 to 1996 in one (centre A) and for 1995 (6 months) in the other (centre B). A total of 954 and 230 toenails and 621 and 190 fingernails, respectively, underwent KOH microscopy and culture studies in each centre. Positive findings were noted in 45 and 65% of the toenails and 44 and 72% of the fingernails, respectively. In the toenails, Candida spp. were found in 22 and 15%, respectively, and in the fingernails, in 77 and 63%, respectively. The most frequent Candida species was C. parapsilosis (39.5% in toenails, 36.7% in fingernails), followed by C. albicans (19.5% in toenails, and 34.4% in fingernails). These results demonstrate a higher frequency of isolation of C. parapsilosis compared with C. albicans in onychomycosis. This might have important therapeutic implications.
Case Report. Successful outcome of invasive nasal sinus zygomycosis in a child with relapsed acute lymphoblastic leukaemia due to liposomal amphotericin B
Authors: Wali Y. A. , Al Lamki Z. , Al Kindi H. , Taqi A. M. , Shah W. , Soliman H. , Zackaria M. , Al Okbi H.
Published: March 2003

Summary.  We report a case of life‐threatening nasal sinus zygomycosis that developed during remission induction therapy for a relapsed acute lymphoblastic leukaemia. The patient was successfully treated with liposomal amphotericin B and granulocyte‐colony stimulating factor followed by surgical reconstruction of the resultant cutaneous defect.
Case report. Onychomycosis due to Microsporum canis
Authors: Romano C. , Paccagnini E. , Pelliccia L.
Published: March 2003

Summary. A case of distal subungual onychomycosis of the big toe due to Microsporum canis is reported in a 69‐year‐old male asthma patient who had been treated with systemic corticosteroids for the last 3 years. The nail infection was contracted from a cat who was a healthy carrier. The patient was treated successfully with intermittent itraconazole therapy.