A survey of enteric bacteria and protozoans in fresh bovine faeces on New Zealand dairy farms

Aims: To determine the counts and/or prevalence in fresh bovine faeces of Escherichia coli, enterococci, Campylobacter, Salmonella, shiga toxin‐producing E. coli (STEC), Giardia and Cryptosporidium, as inputs to numerical models designed to estimate microbial loadings on pasture grazed by cattle in New Zealand. Methods and Results: In each season over one year, samples of freshly deposited bovine faeces were collected from four New Zealand dairy farms (n = 155), and enumerated for E. coli, enterococci, Campylobacter, Giardia and Cryptosporidium. They were also tested for the presence of Salmonella and STEC. The overall median bacterial counts (g^?1 wet weight) were E. coli– 5·9 × 10⁶; enterococci – 1·3 × 10⁴; Campylobacter– 3·9 × 10⁵. All counts were highly variable within and between samplings, and few seasonal or regional patterns emerged. However, mean Campylobacter counts were consistently higher in spring. No Salmonella spp. was detected, and only two samples were positive for STEC. Cryptosporidium and Giardia were isolated from 5·2% and 4·5% of the samples, respectively, yielding low numbers of (oo)cysts (1–25 g^?1 and 1–17 g^?1, respectively). Conclusions: Fresh bovine faeces are a significant source of E. coli, enterococci and Campylobacter on New Zealand pastures, although numbers are likely to vary markedly between faecal samples. Significance and Impact of the Study: The study provides the first significant set of indicator and pathogen counts for one of the largest sources of faecal contamination of natural waters in New Zealand, and will be used to model these inputs.     

Alteration of the menstrual cycle in baboons placed on tethering devices and moved to individual housing--a stress model for a follicular phase defect

BACKGROUND: During an attempt to identify endocrine characteristics in the baboon that would more precisely predict ovulatory status for assisted reproductive techniques, we observed severe alterations in the menstrual cycle length upon introducing an environmental stress. This environmental stress involved moving animals from their baseline gang cage environment to individual indoor caging and placing them on a tethering apparatus. METHODS: Five adult female baboons were followed for changes in sex skin indicative of menstrual cycle timing and move from outdoor gang gages to individual indoor cages during the early follicular phase of their cycle. A tether device including a surgically implanted cannula was then installed to facilitate daily blood draws without sedation. Radioimmuonoassays were performed to monitor serum estradiol levels and lapraroscopic surveillance was used to confirm time of ovulation. RESULTS: Complete data sets were collected from four of the female baboons. In each case, a prolongation of the menstrual cycle was noted either during the cycle during which the females were moved to indoor caging or during the cycle immediately following the move. This prolongation was isolated to the follicular phase of the affected cycle. CONCLUSIONS: We conclude that otherwise normal handling procedures, including movement to new caging, and/or installation of a tether device, can impart a stress effect on reproductively cycling adult female baboons, such that folliculogenesis is delayed.     

Microbial fouling community analysis of the cooling water system of a nuclear test reactor with emphasis on sulphate reducing bacteria

Culture and molecular-based techniques were used to characterize bacterial diversity in the cooling water system of a fast breeder test reactor (FBTR). Techniques were selected for special emphasis on sulphate-reducing bacteria (SRB). Water samples from different locations of the FBTR cooling water system, in addition to biofilm scrapings from carbon steel coupons and a control SRB sample were characterized. Whole genome extraction of the water samples and SRB diversity by group specific primers were analysed using nested PCR and denaturing gradient gel electrophoresis (DGGE). The results of the bacterial assay in the cooling water showed that the total culturable bacteria (TCB) ranged from 10³ to 10⁵ cfu ml^?1; iron-reducing bacteria, 10³ to 10⁵ cfu ml^?1; iron oxidizing bacteria, 10² to 10³ cfu ml^?1 and SRB, 2–29 cfu ml^?1. However, the counts of the various bacterial types in the biofilm sample were 2–3 orders of magnitude higher. SRB diversity by the nested PCR-DGGE approach showed the presence of groups 1, 5 and 6 in the FBTR cooling water system; however, groups 2, 3 and 4 were not detected. The study demonstrated that the PCR protocol influenced the results of the diversity analysis. The paper further discusses the microbiota of the cooling water system and its relevance in biofouling.     

The effect of climate change on the occurrence and prevalence of livestock diseases in Great Britain: a review

There is strong evidence to suggest that climate change has, and will continue to affect the occurrence, distribution and prevalence of livestock diseases in Great Britain (GB). This paper reviews how climate change could affect livestock diseases in GB. Factors influenced by climate change and that could affect livestock diseases include the molecular biology of the pathogen itself; vectors (if any); farming practice and land use; zoological and environmental factors; and the establishment of new microenvironments and microclimates. The interaction of these factors is an important consideration in forecasting how livestock diseases may be affected. Risk assessments should focus on looking for combinations of factors that may be directly affected by climate change, or that may be indirectly affected through changes in human activity, such as land use (e.g. deforestation), transport and movement of animals, intensity of livestock farming and habitat change. A risk assessment framework is proposed, based on modules that accommodate these factors. This framework could be used to screen for the emergence of unexpected disease events.     

A specific marker,pat, for studying the fate of introduced bacteria and their DNA in soil using a combination of detection techniques

A specific eucaryotic DNA marker from Solanum tuberosum cv Bintje (688 bp patatin cDNA fragment) was cloned into the unique HindIII-site of plasmid RP4. RP4:: pat was transferred from Escherichia coli to Pseudomonas fluorescens R2f by filter mating.Homology to pat was not detected in the microbial population of Ede loamy sand soil, nor in that of the rhizosphere of wheat growing in this soil, as evidenced by colony filter hybridization. More sensitive molecular detection techniques like most-probable-number recovery/hybridization analysis, and analysis of total community DNA from soil by polymerase chain reaction (PCR) amplification did not reveal the presence of the pat sequence either.P. fluorescens R2f (RP4:: pat), introduced into sterile soil extract microcosms, initially showed poor survival and plasmid loss, after which the introduced populations grew and stabilized at a level of about Log₁₀ 7 cfu per mL. Between 25 and 50% of the population maintained the plasmid, as evidenced by filter hybridization of colonies from non-selective agar plates using the pat fragment as probe.Introduced R2f (RP4:: pat) could be recovered from soil microcosms using selective plating followed by colony hybridization and MPN recovery/hybridization with the pat probe. The presence of the pat marker always coincided with the presence of the resistance genes on RP4:: pat, indicating pat was an adequate marker of the presence of this plasmid. In addition, it adequately described the population dynamics of the introduced strain in soil, since no loss of the plasmid occurred.Hybridization to pat was also useful to show transfer of plasmid RP4:: pat to a recipient strain in soil; transfer to indigenous bacteria was not detected.Analysis by slot-blot hybridization of total community DNA extracted from inoculated soils indicated about Log₁₀ 6 cfu per g of dry soil were still detectable. Application of the PCR on this DNA indicated pat was detectable at least at a level of Log₁₀ 4 immunofluorescence-detectable cells per g of dry soil. Thus extraction of total community DNA followed by PCR permitted the detection of genetically engineered microorganisms present in soil as non-culturable cells.     

Long-term effect of a simple nest-box on the reproductive efficiency and other life traits of an Aotus lemurinus lemurinus monkey colony: an animal model for malaria research

Background The long‐term effect of a PVC pipe nest‐box on the reproductive efficiency and other life traits of an Aotus monkey‐breeding colony have not been characterized. Methods and Results We analyzed laboratory records of the Gorgas Memorial Institute (GMI) Aotus monkey colony in Panama for the period 1999–2010 and found a 273% increase in the annual mean life births in the following 7 years after the introduction of a PVC pipe nest‐box in 2002, as well as increases in the mean body mass and survival of laboratory‐bred monkeys. Other life traits such as inter‐birth interval, parity, birth sex distribution, mortality, and longevity were also determined. Conclusions The use of a PVC pipe nest‐box significantly improved the reproductive efficiency and other life traits of the GMI Aotus breeding colony.     

A cross-sectional analysis of the prevalence of dental anxiety and its relation to the oral health-related quality of life in patients with dental treatment needs at a university clinic in Switzerland

doi: 10.1111/j.1741‐2358.2011.00465.x A cross‐sectional analysis of the prevalence of dental anxiety and its relation to the oral health‐related quality of life in patients with dental treatment needs at a university clinic in Switzerland Objectives: The aim of this observational, cross‐sectional study was to analyse the relationship between dental anxiety (DA) and health‐related quality of life aspects associated with oral conditions of a population with dental treatment needs in Switzerland. Methods: The measurements of DA were collected by means of two questionnaires, the Dental Anxiety Scale (DAS) and a Visual Analogue Scale (VAS). Oral health‐related quality of life (OHRQoL) was assessed with the Oral Health Impact Profile (OHIP). The sample included 223 patients enrolled at a university clinic which specialises in oral prosthetic rehabilitation and temporomandibular disorders. Of them, 78.9% were at or above the age of 50. Results: No gender or age dependencies were observed. A comparison of answers regarding OHRQoL and DA revealed a significant interdependence (p = 0.0118); highly anxious patients were 3.55 times more likely to suffer from poor quality of life compared with less anxious ones. Conclusion: This cross‐sectional study of mostly elderly patients seeking dental treatment in Switzerland found that increased DA was associated with an impaired OHRQoL. The average DA was slightly higher than the results of other industrialised countries and the average OHRQoL was reduced.     

Validation of MALDI-TOF MS for rapid classification and identification of lactic acid bacteria, with a focus on isolates from traditional fermented foods in Northern Vietnam

Aims: To evaluate the potential use of MALDI-TOF MS for fast and reliable classification and identification of lactic acid bacteria (LAB) from traditional fermented foods. Methods and Results: A total of 119 strains of LAB from fermented meat (nem chua) were analysed with both (GTG)(5) -PCR fingerprinting and MALDI-TOF MS. Cluster analysis of the profiles revealed five species represented by a single isolate both in (GTG)(5) -PCR and in MALDI-TOF MS; five species grouped alike for (GTG)(5) -PCR and for MALDI-TOF MS; however, differences in minimal similarity between the delineated (GTG)(5) -PCR and MALDI-TOF MS clusters could be observed; three species showed more heterogeneity in their MALDI-TOF MS profiles compared to their (GTG)(5) -PCR profiles; two species, each represented by a single MALDI-TOF cluster, were subdivided in the corresponding (GTG)(5) -PCR dendrogram. As proof of the identification potential of MALDI-TOF MS, LAB diversity from one fermented mustard sample was analysed using MALDI- TOF MS. PheS gene sequencing was used for validation. Conclusions: MALDI-TOF MS is a powerful, fast, reliable and cost-effective technique for the identification of LAB associated with the production of fermented foods. Significance and Impact of the Study: Food LAB can be identified using MALDI-TOF MS, and its application could possibly be extended to other food matrices and/or other food-derived micro-organisms.     

A novel episomal shuttle vector for transformation of Cryptococcus neoformans with the ccdB gene as a positive selection marker in bacteria

We report the engineering of a new shuttle vector featuring its episomal maintenance in Cryptococcus neoformans and the lethal Escherichia coli ccdB gene for positive selection in bacteria. Telomere-like sequences from C. neoformans and the STAB fragment confer episomal maintenance to the vector (pPM8) upon transformation in C. neoformans. The vector generated high transformation frequencies and each transformant was estimated to harbor thirty copies of the plasmid. The plasmids recovered in E. coli from the C. neoformans transformants showed no evidence of rearrangement. This construct will be very useful for cloning and studying the regulation of genes in C. neoformans.     

Neuronal ceroid lipofuscinosis type CLN2: A new rationale for the construction of phenotypic subgroups based on a survey of 25 cases in South America

Tripeptidyl-peptidase 1 (TPP1) null or residual activity occurs in neuronal ceroid lipofuscinosis (NCL) with underlying TPP1/CLN2 mutations. A survey of 25 South American CLN2 affected individuals enabled the differentiation of two phenotypes: classical late-infantile and variant juvenile, each in approximately 50% of patients, with residual TPP1 activity occurring in approximately 32%. Each individual was assigned to one of three subgroups: (I) n = 11, null TPP1 activity in leukocytes; (II) n = 8, residual TPP1 activity of 0.60–15.85 nmol/h/mg (nr 110–476); (III) n = 6, activity not measured in leukocytes. Curvilinear bodies (CB) appeared in almost all studied CLN2 subjects; the only exceptions occurred in cases of subgroup II: two individuals had combined CBs/fingerprints (FPs), and one case had pure FPs. There were 15 mutations (4 first published in this paper, 3 previously observed in South America by our group, and 8 previously observed by others). In subgroup I, mutations were either missense or nonsense; in subgroups II and III, mutations prevailed at the non-conserved intronic site, c.887 − 10A>G (intron 7), and to a lesser extent at c.89 + 5G>C (intron 2), in heterozygous combinations. Grouping phenotypically and genetically known individuals on the basis of TPP1 activity supported the concept that residual enzyme activity underlies a protracted disease course. The prevalence of intronic mutations at non-conserved sites in subgroup II individuals indicates that some alternative splicing might allow some residual TPP1 activity.     

Impact of predation on early stages of the armoured catfish Hoplosternum thoracatum (Siluriformes-Callichthyidae) and implications for the syntopic occurrence with other related catfishes in a neotropical multi-predator swamp

This study investigated the role of predators in preventing competitive exclusion among three closely related armoured catfishes (Callichthys callichthys, Hoplosternum littorale and H. thoracatum) that occur synthopically in multi-predator freshwater swamps of Suriname, South America. The potential impact of predation on armoured catfish was determined by combining laboratory measurements of predation rates on five early developmental stages of the armoured catfish H. thoracatum for 24 aquatic predators with field studies of the density of the predators in the swamps. The contribution of a particular predator to the total predation pressure on its prey was determined to a large extent by the density of the predator in the swamp. Seemingly innocuous predators with low or moderate predation rates in the laboratory may be extremely important in the swamps due to their high abundance. Small-sized omnivorous fishes and aquatic invertebrates were major predators of early developmental stages of armoured catfish. Both qualitative and quantitative ontogenetic changes in the predation pressure on armoured catfish were observed. Major predation on eggs, larvae and juveniles of H. thoracatum resulted from a different set of predators in each developmental stage of the prey. In all developmental stages of H. thoracatum the predation pressure involved several predator species and not a single, dominant predator. The potential predation pressure of the 24 predators taken together and the number of predators that were able to prey on H. thoracatum decreased sharply with increasing age (size) of the prey. Even if egg (nest) predation is prevented by the guarding male, the potential impact of the 24 predators on the populations of armoured catfish is large. Predation may account for the high mortality of H. thoracatum observed in the swamps. The high predation pressure on callichthyid catfishes may help to explain the coexistence of three closely related and morphologically quite similar armoured catfishes in Surinamese swamps.     

A novel technique for the measurement of temperature in outflow water from a coastal power plant, with notes on chlorination and phytoplankton determinations

This communication discusses a novel approach for the measurement of temperature in the mixing area of outflow water from Madras Atomic Power Station, located at a coastal site near Tamil Nadu, India. In addition, various environmental aspects such as chlorination, phytoplankton estimation and annual sea surface water temperature have also been addressed in this paper in the light of similar work published earlier from the same locality.