bluuebunny/crossref_metadata_2025 huggingface dataset:
Development of an ELISA system for Detection of Anti‐Anaplasma marginale Antibodies in cattle in Brazil
Authors: Braz C.J. , Ribeiro M.F.B. , Lima J.D. , Passos L.M.F.
Published: March 2003

An ELISA test was developed for detecting antibodies against Anaplasma marginale in bovine sera. Four antigenic preparations were produced from infected red blood cells. Some aliquots of this preparation were stored at ‐70°C with 30% DMSO in phosphate‐buffered saline (PBS) and others were lysed with 0.9% NH4Cl and stored at ‐ 20°C. Typical anaplasmal structures were seen by electron microscopy in the antigenic preparations containing the erythrocytes that had been stored with DMSO. The performance of the ELISA test was evaluated by testing 298 positive serum samples collected from immunized cattle, 39 negative serum samples collected from cattle imported from areas free of A. marginale and 50 samples collected from cattle naturally infected in the field. The test gave a specificity of 94.87% and a sensitivity of 100%.
Scanning Electron Microscopy of Adult Gongylonema pulchrum (Nematoda: Spirurida)
Authors: Naem S. , Seifi H. , Simon G.T.
Published: March 2003

Scanning electron microscopy (SEM) was used to study the surface ultrastructure of adult worms of Gongylonema pulchrum. The anterior end in both sexes was covered by numerous cuticular platelets. There was a pair of lateral cervical papillae. The buccal opening was small and extended in the dorsoventral direction. Around the mouth a cuticular elevation enclosed the labia, and eight papillae were located laterodorsally and lateroventrally. Two large lateral amphids were seen. On the lateral sides of the female's tail, phasmidal apertures were observed. The caudal end of the male was asymmetrically alate and bore 10 pairs of papillae and two phasmidal apertures.
Development of a Differential Multiplex PCR Assay for Equine Herpesvirus 1 and 4 as a Diagnostic Tool
Authors: Carvalho R. , Passos L. M. F. , Martins A. S.
Published: March 2003

In this study, a multiplex polymerase chain reaction (PCR) procedure was developed for differentiation of strains and field isolates of equine herpesvirus type 1 (EHV‐1) and type 4 (EHV‐4). Specific oligonucleotide primers were combined to amplify the thymidine kinase (TK) gene region of EHV‐1 and EHV‐4, which would yield fragments of different lengths for each virus in the same amplification reaction. The specificity of the largest PCR amplicon for EHV‐4 was confirmed by restriction digestion with HindIII. The multiplex PCR proved to be a fast and sensitive method for typing EHV‐1 and EHV‐4 isolates and for detection and differentiation of both viruses in field samples in which infectious virus is no longer available. The sensitivity was improved by combining cycling optimization and visualization of PCR products in ethidium bromide and silver‐stained acrylamide gels.
Flow Cytometry for the Detection of Enterohaemorrhagic Escherichia coli O157 : H7 with Latex Beads Sensitized with Specific Antibody
Authors: Kusunoki H. , Latiful Bari M. , Kita T. , Sugii S. , Uemura T.
Published: March 2003

To detect low concentrations of enterohaemorrhagic Escherichia coli O157 : H7 rapidly, flow cytometry (FCM) was carried out with specific IgG‐sensitized latex beads (IgG‐Lx). It was found that test samples for FCM can be prepared for much shorter periods by culturing E. coli O157 : H7 in trypto‐soya broth at 42°C and by treatment with 0.5 % formalin at 37°C. FCM with IgG‐Lx performed with E. coli O157 : H7 prepared by such a procedure revealed that the lowest number of E. coli O157 : H7 prepared in pure culture detected by FCM was 103/ml. Because similar findings have already been reported by FCM with immunomagnetic beads, FCM with IgG‐Lx is also suggested to be a valuable technique to detect low numbers of E. coli O157 : H7 rapidly in food stuffs.
Species of Flea (Siphonaptera) Infesting Pets and Hedgehogs in Germany
Authors: Visser M. , Rehbein S. , Wiedemann C.
Published: March 2003

The species of flea infesting pets and hedgehogs in Germany were investigated through a survey of small animal practitioners throughout the country who were asked to collect specimens at their veterinary practices. A total of 625 veterinarians/veterinary practices responded and provided 2445 intact and identifiable flea specimens. These fleas originated from 294 dogs (795 fleas), 334 cats (1152 fleas), 76 hedgehogs (481 fleas), five domestic rabbits (10 fleas), one golden hamster (four fleas) and one ferret (three fleas). Dogs were found to be infested with Archaeopsylla erinacei, Chaetopsylla globiceps, Ctenocephalides canis, Ctenocephalides felis, Hystrichopsylla talpae, Nosopsyllus fasciatus, Paraceras melis and Pulex irritans. From cats, Archaeopsylla erinacei, Ceratophyllus gallinae, Ceratophyllus garei, Ctenocephalides felis, Ctenophthalmus assimilis, Hystrichopsylla talpae, Monopsyllus sciurorum, Nosopsyllus fasciatus, Spilopsyllus cuniculi and Typhloceras poppei were collected. In both dogs and cats the most prevalent species were Ctenocephalides felis (78.9% and 91.6%, respectively) and Archaeopsylla erinacei (21.1% and 12.6%, respectively) followed by Ctenocephalides canis in dogs (5.8%) and Hystrichopsylla talpae in cats (1.2%). The fleas isolated from rabbits were Ctenocephalides felis, Hystrichopsylla talpae and Spilopsyllus cuniculi. Nosopsyllus fasciatus and Ctenocephalides felis were recovered from the golden hamster and the ferret, respectively. The hedgehogs were found to be infested with Archaeopsylla erinacei, Ceratophyllus gallinae and Ctenocephalides felis.
Hydatid Echinococcosis (Echinococcus granulosus) in a Laboratory Colony of Pig‐Tailed Macaques (Macaca nemestrina)
Authors: Plesker R. , Bauer C. , Tackmann K. , Dinkel A.
Published: March 2003

Several cases of hydatid echinococcosis were diagnosed in a laboratory colony of 19 pig‐tailed macaques (Macaca nemestrina) at the Paul Ehrlich Institute, Germany. Three hydatid cysts were found in the liver of an euthanized animal. The diagnosis of an Echinococcus granulosus infection was confirmed by histopathology and the results of a specific polymerase chain reaction. The serum of five of 14 other monkeys tested for Echinococcus antibodies using a genus‐specific enzyme‐linked immunosorbent assay (ELISA) was positive or weakly positive; none of the animals, however, showed specific reactions in an E. multilocularis‐specific ELISA. On ultrasonographic examination, alterations in the liver were found in four of the serologically positive monkeys, and two animals showed clinical signs such as progressive anorexia, apathy and icterus. The monkeys had most probably acquired the E. granulosus infection in their breeding colony in Slovenia.
Dynamics of Staphylococcus aureus Infections during Vaccination with an Autogenous Bacterin in Dairy Cattle
Authors: Hoedemaker M. , Korff B. , Edler B. , Emmert M. , Bleckmann E.
Published: March 2003

The effect of an autogenous vaccine against Staphylococcus aureus on S. aureus prevalence and mastitis, as well as on somatic cell count (SCC), was studied in a dairy herd with a high prevalence of S. aureus. The vaccination group (n=35; 22 cows and 13 heifers) and the control group (n=36; 23 cows and 13 heifers) received the vaccine or a placebo, respectively, according to the following protocol: all animals: basic immunization (twice, 3 weeks apart); cows: booster dose at the time of drying off, 5 and 2 weeks before calculated calving date; heifers: booster dose 2 and 5 weeks before calculated calving date. The vaccine or the placebo was administered subcutaneously in the area of the supramammary lymph nodes. Quarter milk samples were collected monthly and subjected to SCC and bacteriological evaluation. At this time, the animals were also checked for signs of clinical mastitis. Non‐clinical S. aureus mastitis diagnoses were based on udder quarter SCC and a positive S. aureus culture. In order to compare the SCC in individual whole milk samples, records from the monthly milk quality testing were evaluated. Cow and udder quarter prevalence of S. aureus intramammary infections calculated for the experimental animals and quarters, respectively, did not differ between groups. However, during the lactation period following the booster dose, the prevalence of S. aureus increased in both groups (P < 0.05). The cumulative incidence of various mastitis diagnoses (clinical, subclinical, latent infection) due to S. aureus on an animal basis did not differ between groups. On an udder quarter basis, the cumulative incidence of subclinical mastitis was higher in vaccinated animals than in control animals (33.8 versus 26.0 %; P < 0.05). This was mainly due to a higher cumulative incidence of subclinical mastitis in vaccinated than control heifers. The SCC in composite milk samples did not differ between groups, but increased as lactation progressed. The herd prevalence of S. aureus differed considerably throughout the study period, but declined consistently to below 10 % at the end of the study period. Recent herd checks revealed a prevalence of S. aureus infections of < 5 %. It is concluded that the autogenous bacterin tested in this study did not have the desired effect on the prevalence of S. aureus infections and mastitis or SCC. The decline in S. aureus prevalence was very probably due to other factors than specific immunization against S. aureus.
Severe Enteric Disease in an Animal Shelter Associated with Dual Infections by Canine Adenovirus Type 1 and Canine Coronavirus
Authors: Pratelli A. , Martella V. , Elia G. , Tempesta M. , Guarda F. , Capucchio M. T. , Carmichael L. E. , Buonavoglia C.
Published: March 2003

An outbreak of dual infection in dogs with canine adenovirus type 1 (CAV‐1) and canine coronavirus (CCV) infection is reported in an animal shelter that comprised approximately 200 adults stray dogs and 30 puppies. Twenty puppies died 7–8 days after the onset of the clinical signs (severe enteritis, leucopoenia, respiratory distress and dehydration). Both CAV‐1 and CCV were isolated from tissue or swab samples. Antibodies to CCV and, at high levels, to CAV‐1 also were detected in several puppies. The principal histological findings were atrophy of small intestinal villi, lymphoid depletion, hepatitis and bronchopneumonia. The persistence of CCV in the faeces, observed by the polymerase chain reaction assay, was longer than previously reported. Results demonstrated the serious consequences which may occur with dual infections by CAV‐1 and CCV in assembled groups of dogs that are housed in poorly managed kennels with inadequate vaccination programmes.
Entropion, Corneal Ulcer and Corneal Haemorrhages in a One‐Humped Camel (Camelus dromedarius)
Authors: Yeruham I. , Van Straten M. , Elad D.
Published: March 2003

An unusual case of entropion, corneal ulcer and corneal haemorrhages in a one‐humped camel (Camelus dromedaries) is described. The most prominent clinical findings were entropion of both eyelids, severe blephrospasm, epiphora, conjunctivitis, conjunctival oedema, mucopurulent conjunctival discharges, hyperaemia, lacrimation and photophobia. Corneal ulcers and corneal haemorrhages were also observed.
Nested Reverse Transcriptase‐Polymerase Chain Reaction for the Detection of Group A Rotaviruses
Authors: ELSCHNER M. , PRUDLO J. , HOTZEL H. , OTTO P. , SACHSE K.
Published: March 2003

Rotaviruses are important pathogens associated with diarrhoeal diseases in almost all species of mammals. In the present study, a nested reverse transcriptase‐polymerase chain reaction (RT‐PCR) for the detection of group A rotaviruses was developed, which is based on a target region in gene segment 6. Rotavirus strains of human, bovine, porcine, canine, feline, equine, and ovine origin were examined. Furthermore several faecal specimens, in which rotavirus had already been detected using other methods than PCR, were included in the study. A nested RT‐PCR product was formed with all strains and faecal samples tested. The detection limit for virus‐containing cell culture supernatant was 3 × 10–2 [50% tissue culture infective dose (TCID50)] by RT‐PCR and 3 × 10–3 TCID50 by nested amplification. In order to examine the influence of the sample matrix on sensitivity, a rotavirus‐negative faecal specimen was spiked with virus‐containing cell culture suspension of the porcine rotavirus OSU. The detection limit of the present PCR procedure was approximately 1.6 × 102 TCID50 per g faeces and could be increased by one order of magnitude using nested PCR. The present method for detection and identification of group A rotaviruses represents a powerful diagnostic tool and was shown to be applicable to rotaviruses of different origin, including human sources.
Escherichia coli Isolates from Young Calves in Bavaria: In Vitro Susceptibilities to 14 Anti‐microbial Agents
Authors: WERCKENTHIN C. , SEIDL S. , RIEDL J. , KIOSSIS E. , WOLF G. , STOLLA R. , KAADEN O.‐R.
Published: March 2003

During the occasional testing of Escherichiacoli from faecal samples of young calves we observed multi‐resistant isolates. Because of the significance of E. coli as an indicator bacterium for resistance trends we tested E. coli populations of young calves over a longer period. Here we present the results of a retrospective study comparing isolates from 1998 to 2000. Moreover, we compared, in a clinical study, the resistance rates of E. coli populations from 67 hospitalized calves both before and after hospitalization (with or without anti‐microbial therapy), and with their anamnestic data of antibiotic usage. The highest resistance rates were found to be more than 80% for tetracyclines, ampicillin, sulfonamide/trimethoprim combinations, and chloramphenicol. A significant increase or decrease over the years was not observed. In analysing the data of hospitalized calves, an increase of resistance to some anti‐microbials had to be registered that seemed to be connected with the selective pressure due to agents used in the clinic. In comparing anamnestic data and resistance rates it became obvious that reliable data are not easily available and that a number of potential anti‐microbial influence factors have to be taken into account.
Patterns ofMycoplasma hyopneumoniaeInfections in Belgian Farrow‐to‐Finish Pig Herds with Diverging Disease‐Course
Authors: Vicca J. , Maes D. , Thermote L. , Peeters J. , Haesebrouck F. , de Kruif A.
Published: March 2003

Patterns ofMycoplasma hyopneumoniae(Mh) infections were investigated in five clinically infected herds and in five herds subclinically infected with Mh. In the clinically infected herds, housing and management conditions were good whereas these conditions were poor in the subclinically infected herds. In each herd, serum antibodies against Mh were detected in pigs of different ages and nasal swabs were taken for Mh detection using nested PCR (nPCR). The percentage of seropositive pigs in the clinically infected herds increased from 8% in pigs of 9 weeks to 52% in pigs of 18 weeks and seroconversion was most shown between 12 and 15 weeks. In the subclinically infected herds, the percentages increased from 2 to 24% and most of the pigs became seropositive between 15 and 18 weeks. The percentage of nPCR positive pigs at 6 weeks was 16 and 0% in the clinically and subclinically infected herds, respectively. The results demonstrate that the seroprevalences were higher in the clinically infected herds and that most of the pigs became infected with Mh at a younger age. It can be concluded that additional factors different from housing and management, like differences among Mh strains, may determine the infection pattern of Mh and the clinical course of the infection.
Transmission ofAnaplasma phagocytophila(Human Granulocytic Ehrlichiosis Agent) in Horses Using Experimentally Infected Ticks (Ixodes scapularis)
Authors: Pusterla N. , Chae J.‐S. , Kimsey R. B. , Berger Pusterla J. , DeRock E. , Dumler J. S. , Madigan J. E.
Published: March 2003

Most human granulocytic ehrlichiosis (HGE) studies carried out in horses use needle inoculation of infected leucocytes or cell cultures. This route of inoculation does not accurately reflect natural infection of the tick‐borne agent. To investigate whether tick transmission influences the course of granulocytic ehrlichiosis in the horse model, experimental transmission through infected laboratory‐rearedIxodes scapularisticks was attempted into two healthy horses. One additional horse served as negative control and was exposed to uninfected ticks. Eleven days after exposure to nymphal or adult ticks infected withAnaplasma phagocytophila(HGE agent) the two horses developed severe clinical and laboratory signs consistent with granulocytic ehrlichiosis. Bacteraemia was determined at various time points in the two horses by observation of morulae within neutrophils and by detection ofA. phagocytophilagenomic DNA by PCR of peripheral blood leucocytes. Further, both horses seroconverted. In contrast the control horse stayed uninfected. The results demonstrate thatA. phagocytophilacan be experimentally transmitted by infected nymphal and adult ticks and that the agent is able to produce a severe disease, similar to naturally occurring cases. Therefore, tick transmission is highly reproducible and can be successfully used in the equine animal model in order to study HGE.
Foetal Protection against Bovine Virus Diarrhoea Virus after Two‐step Vaccination
Authors: Frey H.‐R. , Eicken K. , Grummer B. , Kenklies S. , Oguzoglu T. C. , Moennig V.
Published: March 2003

In order to assess the efficacy of a two‐step vaccination protocol with respect to foetal protection against transplacental infections with bovine virus diarrhoea virus (BVDV) with special attention to BVDV‐2 seronegative heifers were vaccinated with an inactivated BVDV‐1 vaccine and boostered with a modified live BVDV‐1 vaccine after 4 weeks. A second group was left unvaccinated as control. Between days 30 and 120 of pregnancy the heifers of both groups were intranasally challenged with a mixture of BVDV‐1 and ‐2. All heifers of the vaccinated group gave birth to nine clinically healthy, seronegative (precolostral) and BVDV‐free calves. In contrast in the control group four BVDV viraemic underdeveloped calves were born. Additionally, one calf was stillborn and another viraemic calf was not viable and died 2 days after birth. All six calves of the control group were viraemic with BVDV‐2.

This study demonstrated for the first time that two‐step vaccination of breeding cattle with a modified live BVDV vaccine 4 weeks after application of an inactivated BVDV vaccine was capable of providing a foetal protection against transplacental infection with BVDV‐2.
Application of Different Methods for the Diagnosis of Paratuberculosis in a Dairy Cattle Herd in Argentina
Authors: Paolicchi F. A. , Zumarraga M. J. , Gioffre A. , Zamorano P. , Morsella C. , Verna A. , Cataldi A. , Alito A. , Romano M.
Published: March 2003

Paratuberculosis (Ptbc) has a high prevalence in Argentina, that affects dairy and beef cattle. The culture is the gold standard to the diagnosis of the disease.Mycobacterium aviumssp.paratuberculosis (M. paratuberculosis), the aetiological agent, is difficult to isolate and grow in culture. In this study, 24 randomly selected cows of the Fresian breed from a dairy herd with a history of Ptbc were used to evaluate the performance of different diagnostic techniques. These animals did not show clinical signs of the disease. However, another animal from this herd presented evidence of clinical disease at the moment of the present study. This animal was necropsied and one strain ofM. paratuberculosiswas isolated from faeces, lymph nodes and intestine. Serum for indirect absorbed enzyme‐linked immunosorbent assay (ELISA) and agar gel immunodiffusion (AGID) tests and whole blood samples to perform gamma interferon (γIFN) release assays were obtained from each animal. Faeces and milk samples to carry out bacteriological cultures, PCR identification ofM. paratuberculosis, and direct examinations of smears with Ziehl–Neelsen's (ZN) stain were also collected. Tuberculin test with bovine purified protein derivative (PPD) in the caudal fold was performed. The results showed that 10 out of 24 animals (41.6%) were positive to ELISA. Eight strains ofM. paratuberculosiswere isolated, six from faeces, two from milk. Five of the animals that excreted the bacteria through faeces were ELISA‐positive, whereas the excreters through milk were negative to ELISA. No positive samples by AGID were obtained in clinical asymptomatic animals. Seven samples gave positiveγIFN results with avian PPD, but only two of these animals were confirmed with culture. Direct PCR, to detect IS900(M. paratuberculosis) in faeces and milk samples, was negative, but PCR using material taken from faecal and milk cultures gave positive results before visualizing the colonies. No sample was positive by PCR directed to IS6110(M. tuberculosiscomplex). There was not always agreement between isolations and ZN in the studied samples. In conclusion, the absorbed ELISA was useful to detect positive animals and excreters through faeces but not through milk. PCR applied to cultures with incipient development before the visualization of colonies was effective to specifically determine the presence ofM. paratuberculosis. TheγIFN test was not able to detect the most positive animals confirmed by culture. The importance of using ELISA and cultures is emphasized by this study but it is necessary to continue with theγIFN test development for early detection of the disease.
Retrospective study on porcine circovirus type 2 infection in pigs from 1985 to 1997 in Spain
Authors: Rodríguez‐Arrioja G. M. , Segalés J. , Rosell C. , Rovira A. , Pujols J. , Plana‐Durán J. , Domingo M.
Published: March 2003

A retrospective survey was performed to detect lesions of Postweaning multisystemic wasting syndrome (PMWS) and nucleic acid of porcine circovirus type 2 (PCV2) in archived formalin‐fixed, paraffin‐embedded tissues from 189 pigs, and antibodies to this virus in sera of 388 pigs from the Spanish livestock between the years 1985 and 1997. PCV2 nucleic acid was detected byin situhybridization (ISH) in tissues from 78 of 189 (41.3%) examined pigs. Variable amount of viral genome was detected in association with slight to severe microscopic lymphoid lesions consisting of lymphocyte depletion and histiocytic infiltration. The first positive case of PMWS with typical lesions and ISH positive corresponded to a pig necropsied in 1986. Two hundred and eighty‐two of 388 (72.7%) sera were positive by immunoperoxidase monolayer assay. Serological and pathological data of the present study indicate that PCV2 was a enzootic infection in Spain since 1985, suggesting that the introduction of this virus in the livestock occurred previously.
Histological Identification of Macroalgae from Stomach Contents of the Antarctic Fish Notothenia coriiceps Using Semi‐thin Sections
Authors: Iken Katrin , Quartino Maria‐Liliana , Wiencke Christian
Published: March 2003

A histological method is described allowing the identification of macro‐algal fragments from stomach contents of fish. The rhodophytes Porphyra endiviifolium and Neuroglossum ligulatum were determined from the stomach contents of the Antarctic fish Notothenia coriiceps. The results show P. endiviifolium to be a more important part in the diet of this fish than previously expected. On the basis of the depth distribution of the identified algae it is suggested that N. coriiceps can migrate to very shallow waters for feeding.
The Sample Size Necessary to Assess Changes in Fish Biomass – A Reply
Authors: Mouillot David , Culioli Jean‐Michel
Published: March 2003

Abstract. In this article, we compare biomass results for three fish species (Symphodus ocellatus, Serranus scriba and Diplodus annularis) between May 1993 and May 2000 in the Bonifacio Strait Marine Reserve. The data were obtained from seagrass meadows using the visual census method. A randomised procedure was used to test the null hypothesis of biomass equality. To this end, decreasing sample sizes were used for the May 2000 data to determine the sample size necessary to reject the null hypothesis (5 % level). For Symphodus ocellatus, 10 samples (fixed points) were necessary to reject the null hypothesis, whereas 15 samples were required for Diplodus annularis. The null hypothesis was never rejected for Serranus scriba regardless of the sample size considered. These results are discussed, as is the power statistical analysis proposed by Ortiz (2002; this issue, pp. 1 – 9), to determine the optimal sample size necessary to detect perturbation effects in the marine environment. Although the power analysis is certainly a useful method to determine the sampling design in assessment the effect of marine reserves on fish biomass, the magnitude of the marine reserve effect remains problematic in ecology.
Lack of sialidase activity in Candida albicans and Candida glabrata
Authors: Roggentin P. , Krug G. , Schauer R. , Brasch J.
Published: March 2003

Because several micro‐organisms having close contact to animal hosts and man produce sialidase (EC 3.2.1.18) as a tool for adhesion and invasion, we investigated two Candida species for the presence of this enzyme. Two sensitive assays, a fluorometric test with 4‐methylumbelliferyl‐α‐d‐N‐acetylneuraminic acid and a radiometric test with 3H‐labelled sialyllactitol as sialidase substrates, were applied to detect sialidase activity. None of 40 Candida albicans and 10 C. glabrata strains grown in three different media exhibited sialidase activity, but the positive control Ophiostoma stenoceras produced sialidase under comparable conditions. Our surprising negative findings are divergent from an earlier positive report, which may be due to strain selection or bacterial contamination. These results indicate that sialidase is probably of no relevance in cutaneous or mucosal candidoses.
A new, semi‐automated technique for quantification of Candida adherence to denture acrylic surfaces
Authors: Panagoda  , Samaranayake
Published: March 2003

Adherence of Candida species to various host surfaces is an important prerequisite for colonization and pathogenesis. Researchers have problems in quantifying candidal adherence to plastic surfaces (used for fabrication of prostheses) due to coaggregation of adherent yeasts and to the laborious, time‐consuming method of microscopic quantification. We describe here, a semi‐automated image analysis system which is superior to conventional visual light microscopic techniques of quantifying yeasts attached to solid substrates such as acrylic. The results demonstrated a significant positive correlation between the number of cells quantified by traditional light microscopy and the corresponding area obtained by the semi‐automated image analysis system (r=0.83, P<0.02). The new system is simple, less laborious and is particularly useful when the adherent attributes of a large battery of yeasts are characterized.
Fungal surveillance cultures during antifungal prophylaxis with itraconazole in neutropenic patients with acute leukaemia
Authors: Glasmacher A. , Hahn C. , Molitor E. , Sauerbruch T. , Schmidt‐Wolf I. G. H. , Marklein G.
Published: March 2003

Fungal colonization has been associated with an increased rate of invasive fungal infections in neutropenic patients. This study evaluates weekly fungal surveillance cultures from the oropharyngeal and perianal space as well as other suspected sites in 219 courses of myelosuppressive chemotherapy with itraconazole antifungal prophylaxis in 116 neutropenic patients with acute leukaemia. Itraconazole was given from the start of chemotherapy in one of six different dosing regimens. Fungal colonization occurred in 68 (31%) of courses, which was lower than in a historical control group without prophylaxis (53%, P=0.004). Twenty‐six per cent of these 116 isolates had a growth rate of more than 50 colony forming units (CFU) per culture. Candida glabrata (51%), Candida albicans (18%) and Candida krusei (4%) were the most frequently isolated species. Higher median itraconazole trough concentrations were associated with a lower growth rate in the cultures (≤50 CFU/culture versus>50 CFU/culture): 710 (430–1180) ng ml−1 versus 900 (560–1650) ng ml−1 (P=0.015). The use of itraconazole solution—compared with capsules—led to a reduced growth rate (P=0.035). In conclusion, compared with historical controls itraconazole antifungal prophylaxis reduces the incidence and the extent of fungal colonization during neutropenia in patients with acute leukaemia.
Studies on the influence of combined administration of ochratoxin A, fumonisin B1, deoxynivalenol and T2 toxin on immune and defence reactions in weaner pigs
Authors: Müller G. , Kielstein P. , Rosner H. , Berndt Angela , Heller M. , Köhler Heike
Published: March 2003

In weaners, combined administration of fumonisin, deoxynivalenol and T2 together with ochratoxin A in quantities expected to be present in feeds of central European origin resulted, as a rule, in changes identical to those observed after single administration of ochratoxin A. Such reactions were partly compensated. Synergistic amplification of immunosuppressive changes due to the simultaneous intake of these mycotoxins in low concentrations is not to be expected.
Dermatophytosis due to Trichophyton violaceum in Tuscany from 1985 to 1997
Authors: Romano Clara , Massai  , Difonzo Elisa M.
Published: March 2003

We describe 45 cases of dermatophytosis due to Trichophyton violaceum diagnosed in Tuscany from 1985 to 1997. Diagnosis was based on direct microscope mycological examination and culture. Nine of the 45 patients were Italian, eight of whom were born and resident in Tuscany; the others were Africans who contracted the infection in their country of origin. The first case in a non‐European was observed in 1985, but since 1991 isolation from foreigners has become more frequent. The increase in cases of infection by T. violaceum in our region is certainly linked to immigration. The cases of epidermomycosis in Tuscans suggests that the fungus has already become autochthonous.
Incidence ofCandidain psoriasis – a study on the fungal flora of psoriatic patients
Authors: Waldman A. , Gilhar A. , Duek L. , Berdicevsky I.
Published: March 2003

Summary. The presence ofCandida albicansand otherCandidaspecies in saliva and faeces of 50 psoriatic patients compared with a control group of 50 healthy donors was examined quantitatively. The quantity ofCandidain saliva and faeces of the psoriatics proved to be significantly higher than in the controls.Candidawas detected in 78% of the saliva samples of the psoriatics but in only 50% of the controls, and in the faeces samples in 72% of the psoriatics, but in only 46% of the controls. Qualitative analysis revealed a predominance ofCandida albicans(saliva, 77%; faeces, 64%) andCandida rugosa(saliva, 28%; faeces, 28%). We did not find a correlation between the severity of the psoriasis according to the Psoriasis Area and Severity Index and the amount ofCandidain the saliva or in the faeces. Our results reinforce the hypothesis thatC. albicansis one of the triggers to both exacerbation and persistence of psoriasis. We propose that in psoriatics with a significant quantity ofCandidain faeces, an antifungal treatment should be considered as an adjuvant treatment of psoriasis.
The aetiological agents of tinea capitis in Zaragoza (Spain)
Authors: Rubio‐Calvo C. , Gil‐Tomas J. , Rezusta‐Lopez A. , Benito‐Ruesca R.
Published: March 2003

Summary. Between 1977 and 1997, 190 cases of tinea capitis were observed. The age groups most commonly infected were 1–10 years old. The following dermatophytes were isolated: Microsporum canis (119), Trichophyton mentagrophytes (57), Trichophyton tonsurans (seven), Trichophyton verrucosum (three), Trichophyton violaceum (one), Trichophyton schoenleinii (one), Trichophyton soudanense (one) and Microsporum audouinii (one).
Dermatophytoses in Prague, Czech Republic, between 1987 and 1998
Authors: Kuklová I. , Kuĉerová H.
Published: March 2003

Summary.  Over a 12‐year period, from 1987 to 1998, the spectrum of aetiological agents isolated from 11 208 patients (6265 females and 4943 males) suspected of having dermatomycosis was analysed in the mycological laboratory of the Department of Dermatology, Charles University, Prague. The most frequently examined locations were toenails (34.9%), feet (15.6%), fingernails (12.8%), toe webs (11%), trunk (10%) and hands (8.7%). Dermatophytes were isolated from 5605 (30.2%) of all 18 528 samples examined. Among dermatophytes, the most common infectious agent was Trichophyton rubrum (90.2%) followed by Trichophyton mentagrophytes (6.6%), Microsporum canis (1.8%) and Epidermophyton floccosum (1.4%). Trichophyton verrucosum, Microsporum persicolor and Microsporum gypseum were rare. During this 12‐year period the pattern of aetiological agents of dermatomycoses in Prague was relatively stable.
Case Report. First isolation of Trichosporon domesticum from a cat
Authors: Sakamoto Y. , Kano R. , Nakamura Y. , Watanabe S. , Kamata H. , Fukuda Y. , Hasegawa A.
Published: March 2003

Summary.  The present study deals with the first isolation of Trichosporon domesticum from the urine of a cat with chronic cystitis. A 5‐year‐old domestic short hair cat weighing 5.6 kg was referred to an animal hospital with the chief complaint of haematuria having persisted for 3 weeks. The isolate was identified as T. domesticum, confirming the result of mycological examination by molecular analysis.
Influence of buffered propionic acid on the development of micro‐organisms in hay
Authors: Reboux G. , Dalphin J.‐C. , Polio J.‐C. , Millon L. , Baverel J. , Martinez J. , Girard L. , Piarroux R.
Published: March 2003

Summary.  We tested the benefit of using buffered propionic acid (BPA) as a means of preventing farmer's lung disease (FLD). BPA, a new formulation of propionic acid, a hay preservative with no deleterious effect on farm machinery or cattle, reduces the development of micro‐organisms in hay. Twenty pairs of round bales were analysed for concentration of micro‐organisms measured in the winter following hay treatment. Each pair included one untreated bale and one bale treated with BPA during haymaking. Our results showed the following decreases in concentration in treated bales: total fungal species, 40% (P < 0.05); Eurotium amstelodami (the main species found), 65% (P < 0.01); and thermophilic actinomycetes, 60% (not significant), respectively. We conclude that BPA could be used to prevent FLD.
Histopathologic and histochemical characterization of calcified structures in hamsters inoculated with Paracoccidioides brasiliensis
Authors: Essayag S. M. , Landaeta M. E. , Hartung C. , Magaldi S. , Spencer L. , Suárez R. , García F. , Pérez E.
Published: March 2003

Summary. In pathogenicity studies of 31 Paracoccidioides brasiliensis isolates preserved using Castellani's method we intraperitoneally inoculated 104 young adult hamsters and found laminated concentric structures and calcified appearance that resembled Schaumann bodies, in 43 of them, especially in animals with apparently good condition. We characterized these structures histologically and histochemically using different stains (PAS, Grocott, haematoxylin‐eosin, Von Kossa). The Von Kossa staining revealed calcium in these structures. Similar structures have been described in patients with sarcoidosis and also in hamsters inoculated with P. brasiliensis. We found no correlation between the presence of these calcifications and serum calcium levels.
Identification of primary trisomics and other aneuploids in foxtail millet
Authors: Wang R. , Gao J. , Liang G. H.
Published: March 2003

A complete set of nine primary trisomics (2n+ 1) for cv.‘Yugu No. 1’of foxtail millet, Setaria italica (L.) Beauv. (2n= 2x= 18), was identified cytologically from progenies derived from crosses between autotriploids(2n= 2x= 27) and their diploid counterparts. Five autotriploid plants were identified from 2100 seedlings derived from 4x‐2x crosses; the reciprocal crosses (2x‐4x) failed to produce autotriploids. Autotriploids grew vigorously and were morphologically very similar to diploids. Theprimary trisomics (2n= 2x= 19) constituted ≅32.5% of the total progeny from the 3x‐2x crosses, whereas 59.8% of the descendants were aneuploids with chromosome numbers ranging from 20 (double trisomics and tetrasomics) to 37 (2n= 4x+ 1; or autotetraploid with one additional chromosome). The nine primary trisomics identified were self‐fertile; seven had characteristic morphology, whereas trisomics VIII and IX resembled the disomics. The seed set for trisomic V was the lowest (20%), and trisomic VIII the highest (74%). Other aneuploids with 20 or more somatic chromosomes were either self‐sterile or partially fertile with various, but low, levels of seed set. Each of the primary trisomics showed its unique transmission rate when self‐pollinated; trisomic IX had the highest (45.8%), whereas trisomic V had the lowest (19.6%) transmission rate.
Inheritance of the resistance to Leptosphaeria maculans of Brassica nigra and B. juncea in near‐isogenic lines of B. napus
Authors: Dixelius C.
Published: March 2003

The inheritance of resistance to Leptosphaeria maculans was studied in near‐isogenic lines derived from asymmetric somatic hybrids between Brassica napus+Brassica nigra and Brassica napus+Brassica juncea, respectively. The hybrids had been backcrossed to B. napus for seven generations before the genetic segregation of the blackleg resistance was determined. The results of the inheritance studies suggested that one single dominant allele controls the resistance in the Brassica napojuncea line, whereas two independent dominant loci were found in the Brassica naponigra line. Total leaf DNA from the near‐isogenic lines was isolated and 89 loci were detected by hybridization to 66 restriction fragment length polymorphism (RFLP) markers previously mapped in the B. nigra genome. Out of the 89 loci, eight loci were detected in the B. naponigra line and six were found in the B. napojuncea line. RFLP markers co‐segregating with blackleg resistance in adult leaves were also found. Two markers associated with linkage group 5 and 8, respectively, of the B genome were found in the B. naponigra line and one marker was associated with linkage group 2 in the B. napojuncea line.
Comparison of isoenzymes in Prunus avium separated by two different electrophoretic techniques
Authors: Pashkoulov D. T. , Tobutt K. R. , BoŠković R.
Published: March 2003

Isoenzyme variation for seven systems revealed by two different electrophoretic procedures was compared in Prunus avium. Fourteen cultivars and 14 wild selections were analysed for acid phosphatase (ACP), isocitrate dehydrogenase (IDH), leucine aminopeptidase (LAP), malate dehydrogenase (MDH), phosphoglucomutase (PGM), shikimate dehydrogenase (SKD) and superoxide dismutase (SOD). Extracts were separated by isoelectric focusing (IEF) and by polyacrylamide gel electrophoresis (PAGE). For the eight loci that had been described previously in these enzyme systems on the basis of IEF analysis, we compared the variation revealed with IEF and PAGE. Similar variation was revealed for Acp‐1 and Pgm‐1, and the alleles revealed by PAGE could be identified directly with those reported for IEF. For Lap‐1, Mdh‐1 and Skd‐1, variation was seen with IEF but not with PAGE. For Mdh‐2, PAGE revealed additional variation not revealed by IEF. For Idh‐1, different patterns of variation were revealed by PAGE and IEF, and both procedures would be needed to genotype cherry accessions. We were unable to detect variation corresponding to that reported previously for Sod‐1 with either technique. The implications of these findings for allele labelling, for studies of genetic diversity and for linkage analysis are discussed.
A chlorophyll‐reduced seedling mutant in oilseed rape, Brassica napus, for utilization in F1 hybrid production
Authors: Zhao Y. , Wang M. L. , Zhang Y. Z. , Du L. F. , Pan T.
Published: March 2003

A mutant chlorophyll‐reduced (Cr) seedling with yellow‐green cotyledons and leaves was obtained from the Brassica napus inbred line 3529 induced by fast neutron and diethyl sulphate (DES). Genetic analysis revealed that the Cr seedling marker trait was controlled by a pair of recessive genes. A randomized complete block design was used to evaluate its agronomic performance. Results from 2 years of tests indicated that the seed yield of Cr lines was significantly lower than that of normal green plants; however, when the Cr gene was in the heterozygous condition, no deleterious effects on yield characteristics and disease resistance were observed. The Cr seedling marker trait was introduced into male‐sterile lines, and Cr male‐sterile lines revealed the same superior combining ability as normal chlorophyll (Nc) lines. The Cr trait can therefore be used as a marker to produce hybrid seed.
Regular segregation of four recessive marker genes among maternal haploids in maize
Authors: Chalyk S. T. , Chebotar O. D.
Published: March 2003

The objective of the study was to investigate whether a population of maternal haploid plants represents a random gametic array. Four inbred lines of maize (A619, MK01, 092 and 19‐3‐3) were used in the present study. These were crossed with line TO carrying four recessive genes: brown midrib (bm2), liguleless (Ig1), white endosperm (g1) and golden plant (g1). Maternal haploids were produced from the F 1 hybrids, using a haploid‐inducing pollinator line. Segregation of haploids for the marker genes was estimated under field conditions. The observed segregation data agreed with the expected 1:1 ratio. It is concluded that no segregation distortion occurred during the production of maternal haploids.
Genetic analysis of resistance to green leafhopper, Nephotettix virescens (Distant), in three varieties of rice
Authors: Angeles E. R. , Khush G. S.
Published: March 2003

The genetics of resistance to green leafhopper, Nephotettix virescens (Distant), in rice varieties ‘IR36’ and ‘Maddai Karuppan’ and breeding line ‘IR20965‐11‐3‐3’ was studied. The reactions of F1 hybrids, F2 populations and F3 lines from the crosses of test varieties with the susceptible variety ‘TN1’ revealed that resistance in ‘IR36’ and ‘Maddai Karuppan’, is governed by single recessive genes while resistance in ‘IR20965‐11‐3‐3’ is controlled by a single dominant gene. Allele tests with the known genes for resistance to green leafhopper revealed that the recessive gene of ‘IR36’ is different from and inherited independently of Glh1, Glh2, Glh3, Glh4, Glh5, Glh8 and Glh9t. This gene is designated as glh10t. The recessive gene of ‘Maddai Karuppan’ and the dominant gene of ‘IR20965‐11‐3‐3’ are also non‐allelic to Glh1, Glh2, Glh3, Glh4, Glh5 and Glh8t. Thus, the dominant gene of IR20965‐11‐3‐3 is designated as Glh11t. The allelic relationships of the recessive gene of ‘Maddai Karuppan’ with glh8 and glh10t should be investigated.
Resistance to Fusarium head blight and deoxynivalenol accumulation in wheat
Authors: Bai G.‐H. , Plattner R. , Desjardins A. , Kolb F. , McIntosh R. A.
Published: March 2003

Fusarium head blight (FHB), caused by Fusarium graminearum Schwabe (telomorph =Gibberella zeae (Schw.)), is an important wheat disease world‐wide. Production of deoxynivalenol (DON) by F. graminearum in infected wheat grain is detrimental to livestock and is also a safety concern in human foods. An international collection of 116 wheat lines was evaluated for FHB resistance and concentration of DON in grain. Plants were inoculated with mixed isolates of F. graminearum in the greenhouse by injecting conidia into a single spikelet of each spike and in the field by scattering F. graminearum‐infected wheat kernels on the soil surface. FHB symptoms were evaluated by visual inspection in both the greenhouse and field, and DON was analysed by HPLC. Significant differences in FHB ratings and DON levels were observed among cultivars. In the greenhouse test, visual symptoms varied from no spread of FHB from the inoculated spikelet to spread throughout the spike, and DON levels ranged from trace levels to 283 mg/kg. In the field test, DON ranged from 2.8 to 52 mg/kg. The greenhouse test identified 16 wheat lines from various origins that accumulated less than 2 mg/kg DON. These lines may be useful as sources for breeding wheat cultivars with lower DON levels. Correlation coefficients were significant between FHB symptom ratings, seed quality traits, and DON levels. Thus, the percentage of scabbed spikelets and kernels can be generally used to predict DON levels in harvested wheat grain. In breeding programmes, selection for plants having few scabbed spikelets and scabbed kernels is most likely to result in low DON levels.
Effect of germination, seed abrasion and seed size on polyphenol oxidase assay activity in wheat
Authors: Demeke T. , Chang H.‐G. , Morris C. F.
Published: March 2003

Polyphenol oxidase (PPO) is one of the major enzymes responsible for browning of wheat food products. Wheat cultivars differ in PPO activity and plant breeders wish to select germplasm and cultivars with low PPO activities. Prior to harvest, seeds may be exposed to conditions conducive to preharvest sprouting and mechanical damage may occur during harvesting and subsequent handling. Sprouting, mechanical damage and seed size may affect PPO assay activity and therefore impinge upon the breeder's ability to accurately select low PPO genotypes. The effects of incipient germination, mechanical abrasion and seed size on PPO assay activity were investigated using the cultivars ‘Klasic’(hard white spring, high PPO), ‘Penawawa’(soft white spring, high PPO) and ‘ID377s’(hard white spring, low PPO). The influence of embryo vs. distal portions of the seed on PPO assay activity was also examined. PPO activity was assayed with constant mixing using L‐3,4‐dihydroxyphenyl alanine (L‐DOPA) substrate at pH 6.5 at room temperature; absorbance of the reaction solution was measured at 475 nm. There was no change in PPO assay activity for seeds imbibed in water for up to 8 h. PPO assay activity increased in seeds imbibed from 8 to 16 h, and then gradually declined with increasing imbibing time. PPO assay activity initially increased as a result of mechanical abrasion but then gradually declined with increasing abrasion time. Large seeds had a higher total PPO assay activity than small seeds, but the difference in activity was not proportional to seed weight. There was only a slight difference in PPO assay activity between half‐seeds with or without the embryo. Overall, PPO assay activity in dry seeds was stable and exposure of seeds to injury and moisture did not make any significant change in PPO assay activity among the cultivars or their relative rankings.
Two major genes for kernel weight in rye
Authors: Wricke G.
Published: March 2003

Previously, kernel weight in rye was considered mostly as a quantitatively inherited trait. F1 plants from parents showing a great difference in kernel weight were selfed and the segregating F2 population was analysed with molecular markers. Two simple sequence repeats markers, one on chromosome 5 and the other on chromosome 7, were found which allowed the genetic analysis of two major genes acting in a complementary way. The analysis of major genes is an advantage in breeding for this important economic trait. In addition it can provide more insight into the genetic structure of this character. This is, in turn, also a prerequisite for future investigations of this trait at the molecular level.