Evidence that host ecology drives first intermediate host use in the Didymozoidae (Trematoda: Hemiuroidea): an asexual infection in a vermetid (Gastropoda).

The Didymozoidae (Trematoda: Hemiuroidea) is among the most speciose trematode families, known from a wide range of marine teleost fishes. Despite their richness, however, didymozoid life cycles are unusually poorly known; only two first intermediate hosts are known, a marine bivalve (Anadara trapezia) and a pelagic gastropod (Firoloida desmarestia). This study uses multi-locus molecular sequence data to identify a novel first intermediate host for the family, a sessile gastropod of the genus Thylacodes Guettard (Vermetidae). The didymozoid infection is not identified to species but, based on molecular phylogenetic analyses, it is close to Saccularina magnacetabula Louvard et al., 2022, which uses a bivalve as a first intermediate host. The distribution of known first intermediate hosts of didymozoids (a bivalve, a holoplanktonic gastropod and a sessile gastropod that feeds with the use of mucus nets) suggests that first intermediate host use within the Didymozoidae has been opportunistically driven by the trophic ecology of potential mollusc hosts and has involved significant host-switching events.

Factors associated with fetal losses in ewe lambs on a New Zealand sheep farm.

CASE HISTORY: As part of a production study of ewe lambs on a large farm in the Waikato region of New Zealand in 2011, pregnancy diagnosis was undertaken twice by trans-abdominal ultrasonography at 68-103 and 97-132 days of gestation. At the second pregnancy diagnosis 257/3,790 (6.8%) ewe lambs had evidence of non-viable fetuses or absence of a pregnancy that was present at the previous pregnancy diagnosis (fetal loss). LABORATORY FINDINGS: Serum antibody titres for Leptospira interrogans serovar Pomona appeared generally higher in 10 ewe lambs with fetal loss compared with 10 that were still pregnant. Histopathological investigation was not able to confirm that fetal loss was associated with leptospial infection. EPIDEMIOLOGICAL INVESTIGATION: In the 2012-born cohort of ewe lambs 443 were vaccinated with a bivalent leptospirosis vaccine and 882 unvaccinated. Serum was collected from 124 non-vaccinated ewe lambs between January and December 2013 for measurement of antibodies to Leptospira serovar Pomona and L. borgpetersenii serovar Hardjo-bovis using a microscopic agglutination test (MAT). Less than 5% of these ewe lambs were seropositive until May, but by August 85% and 48% of animals were seropositive to Leptospira serovars Hardjo-bovis and Pomona, respectively. Fetal loss in non-vaccinated ewe lambs was 78/882 (9%) compared with 23/443 (5%) in vaccinated ewe lambs. Combined data from the 2011- and 2012-born ewe lambs (n=5,115) were analysed using a logistic regression model and fetal loss as the dependent variable. In the final model fetal loss was associated with pre-mating bodyweight (p=0.003), weight change from pre-mating to initial pregnancy diagnosis (p<0.001), year born and leptospirosis vaccination status (p=0.013). Amongst the serologically monitored ewe lambs, there were associations between fetal loss and being seropositive to Leptospira serovar Pomona using titre cut-points of 1:48 and 1:768 (p<0.001). CLINICAL RELEVANCE: Low pre-mating weight and/or low weight gain from mating to pregnancy diagnosis was associated with increased fetal loss, emphasising the importance of ewe lambs achieving target pre-mating weights and liveweight gains during pregnancy. Infection with Leptospira serovar Pomona was associated with fetal loss in the 2012-born cohort and the possibility of infection with this serovar should be considered when investigating cases of fetal loss.

The effects of pasture availability for twin- and triplet-bearing ewes in mid and late pregnancy on ewe and lamb behaviour 12 to 24 h after birth.

This experiment was conducted to evaluate the effect of offering ewes two different feeding levels, during mid and late pregnancy, on ewe and lamb behaviour 12 to 24 h after birth. Romney ewes, bearing twin (n = 80) or triplet foetuses (n = 56), were allocated to a pasture sward height of 2 or 4 cm between 70 and 107 days of pregnancy. In late pregnancy (day 107 to 147), half of the ewes were reallocated the alternate sward height, which produced four treatments: 2-2, 2-4, 4-2 and 4-4. Ewes were weighed on days 65, 92, 107 and 130 of pregnancy and lamb live weights were recorded 12 to 24 h after birth. Twelve to 24 h after birth the maternal behaviour score (MBS) of the ewes were determined, whilst their lambs were tagged. After the lambs were released, the behaviour of each ewe and her lambs was observed for 5 min. Ewe treatment and litter size had no effect on ewe MBS. However, as MBS increased (ewes stayed closer to lambs during tagging), ewes bleated less in a high-pitch and were quicker to make contact with their lamb. During the observation period, ewes in the 4-4 treatment had a greater percentage of their bleats in a low pitch (P < 0.05) than ewes in the 2-2 and 4-2 treatment (61.3% v. 41.3% and 38.8% low bleats, respectively) and more lambs born to 4-4 ewes (95%) bleated than lambs born to 2-2 ewes (84%; P < 0.05). However, lambs born to ewes in the 2-2 treatment bleated earlier than lambs in all other treatments (P < 0.05). Lambs born to 4-4 ewes were less likely (P < 0.05) to move towards their dam in order to make contact than lambs born to 2-2 or 4-2 ewes (3.1% v. 16.9% and 16.7%, respectively). These findings suggest that under the conditions of the present study, ewe nutrition had little effect on maternal behaviour. However, lambs born to ewes offered 2 cm pasture sward heights during mid and/or late pregnancy (2-2, 2-4 and 4-2 treatments) displayed behaviour that demonstrated greater 'need' whereas lambs born to ewes offered 4 cm during mid and late pregnancy sought less attention from their dam.

The effect of different types of stressors during mid- and late pregnancy on lamb weight and body size at birth.

Mid-pregnancy shearing has consistently been shown to increase lamb birth weight, which can lead to an increase in lamb survival rates. However, shearing ewes during the winter months and under outdoor pastoral farming conditions can expose the recently shorn ewe to a greater risk of hypothermia. The aim of this study was to determine if exposure of ewes to repeated stressors, in mid- and late pregnancy, would result in an increase in lamb birth weight. This information may assist in the elucidation of the mechanism for the birth weight response to mid-pregnancy shearing, which in turn could assist in the design of management options to increase lamb birth weight without placing the ewe at risk. One hundred and forty-four twin-bearing Romney ewes were allocated to one of six mid-pregnancy treatments: control, isolation on 2 or 10 occasions, sham-shearing on 10 occasions, intramuscular cortisol injection on 10 occasions or shearing. Isolation, sham-shearing and cortisol treatments were conducted twice a week beginning, on average, day 74 of pregnancy and shearing occurred on day 76. During pregnancy, ewe treatment had no effect on ewe live weight. However, average ewe body condition scores were higher in the shorn group than in the sham-shorn or cortisol groups (P < 0.05). Intramuscular injections of cortisol had a greater effect on ewe plasma cortisol concentrations than all other treatments (P < 0.05). Shearing produced a greater plasma cortisol response than isolation × 10 and sham-shearing (P < 0.05). Ewe plasma cortisol responses decreased during the 5 weeks of isolation and sham-shearing but cortisol injections produced a greater response during the fifth treatment than the first or ninth treatments (P < 0.05). Lambs born to shorn ewes were heavier and had a longer crown rump, forelimb and hind limb lengths than all other lambs (P < 0.05). In addition, lambs born to ewes in the cortisol treatment were lighter than lambs born to control, isolation × 2, isolation × 10 and shorn ewes (P < 0.05). The plasma cortisol concentrations observed for ewes injected with cortisol were far greater than those observed in all other groups, which is likely to explain the low birth weights of lambs born to ewes in that group. These results indicate that the mechanism by which mid-pregnancy shearing increases lamb birth weight is unlikely to be repeated stressors.

A new multiple regression model to identify multi-family houses with a high prevalence of sick building symptoms "SBS", within the healthy sustainable house study in Stockholm (3H).

PURPOSES: The aim was to develop a new model to identify residential buildings with higher frequencies of "SBS" than expected, "risk buildings". METHODS: In 2005, 481 multi-family buildings with 10,506 dwellings in Stockholm were studied by a new stratified random sampling. A standardised self-administered questionnaire was used to assess "SBS", atopy and personal factors. The response rate was 73%. Statistical analysis was performed by multiple logistic regressions. RESULTS: Dwellers owning their building reported less "SBS" than those renting. There was a strong relationship between socio-economic factors and ownership. The regression model, ended up with high explanatory values for age, gender, atopy and ownership. Applying our model, 9% of all residential buildings in Stockholm were classified as "risk buildings" with the highest proportion in houses built 1961-1975 (26%) and lowest in houses built 1985-1990 (4%). CONCLUSION: To identify "risk buildings", it is necessary to adjust for ownership and population characteristics.

How do land-based salmonid farms affect stream ecology?

Increasing research is highlighting the fact that streams provide crucial ecosystem services through the biogeochemical and ecological processes they sustain. Freshwater land-based salmonid farms commonly discharge their effluents into low order, headwater streams, partly due to the fact that adequate freshwater resources for production are commonly found in undisturbed areas. We review the effects of salmonid farm effluents on different biological components of stream ecosystems. Relevant considerations related to the temporal and spatial scales of effluent discharge and ecological effects are discussed. These highlight the need to characterize the patterns of stressor discharge when assessing environmental impacts and designing ecological effects studies. The potential role of multiple stressors in disrupting ecosystem structure and function is discussed with an emphasis on aquaculture veterinary medicines. Further research on the effects of veterinary medicines using relevant exposure scenarios would significantly contribute to our understanding of their impact in relation to other effluent stressors.

Sucking behaviour of hand-reared newborn dairy calves.

AIM: To determine the feeding ability of calves during the first 4 days after birth. METHODS: The sucking behaviour of 171 dairy calves, fed from individual bottles during the first 4 days after birth, was evaluated by quantifying the volume of colostrum consumed, the duration of feeding, and speed of drinking. Calves had been separated from their mothers and brought into a rearing shed in the morning, when < 24 h of age, and were offered 2 L colostrum from a bottle in the afternoon of the same day, and twice daily thereafter. RESULTS: Newborn calves became efficient suckers from bottles within 24 h of removal from their dams (collection). On the day after birth, 95% of calves drank the 2 L of colostrum offered during the afternoon feed, and by Day 4 virtually all calves (99%) drank this amount. Calves that had inadequate colostrum from their dam were more likely to drink all 2 L offered after collection, but a small number of calves that had not had colostrum from their dams and drank < 500 ml at the afternoon feed following collection were likely not to drink 2 L on the following days. However, calves that did not drink all 2 L on Day 1 were not disadvantaged in comparison to those that did, in that they were equally likely to drink 2 L on Day 4. CONCLUSIONS: The majority of calves became efficient drinkers within 48 hours of birth, but a small number of slow feeders may need individual feeding at least up to 4 days after birth.

Identification and enumeration of oleic acid and linoleic acid hydrating bacteria in the rumen of sheep and cows.

The diversity and population densities of facultative anaerobic bacteria with the capacity to hydrate oleic acid and linoleic acid in the rumen of sheep and dairy cows were determined. The screening of representative colonies, from rumen fluid plated aerobically on a range of agar media, revealed that sheep rumen fluid contained hydration-positive strains of Streptococcus, Staphylococcus, Enterococcus, Lactobacillus and Pediococcus, whereas cow rumen fluid contained hydration-positive strains of Streptococcus, Lactobacillus and Staphylococcus. Mean counts of facultative anaerobic bacteria in sheep and cattle rumen were log10 7.29 and log10 6.40, respectively, and were independent of diet. Approximately 56% of facultative anaerobic bacteria were able to hydrate oleic and/or linoleic acid in anaerobic broth culture. For both sheep and cows, the most numerous hydration-positive isolates were strains of Strep. bovis. The results, which are the first to show that pediococci have the capacity to hydrate unsaturated fatty acids, suggest that lactic acid bacteria are the major unsaturated fatty acid hydrating bacteria in the rumen.

Construct validity of a work environment impact scale.

This study examined the construct validity and the internal consistency of a newly developed assessment, the Work Environment Impact Scale (WEIS). After administration to 20 individuals with psychiatric disabilities, Rasch analysis was utilized to scrutinize the data. For this study, criteria for determining unexpected person/item responses were based on the following criteria: MNSQ > 1.3 and $\underline{t}>2.0$. Persons/items with MNSQ < 0.7 and $\underline{t}<-2.0$ were examined to enhance instrument precision but were not considered misfit. Results suggest that the WEIS is an appropriate and valid instrument to utilize with workers who have psychiatric disabilities. Overall, the items appeared to match the worker's need for performance, satisfaction, and well-being as the mean of persons measure is 0.30 ± 0.49 logits more than the mean of items measured. In addition the hierarchical order of items is consistent with literature identifying environmental press and affordance for workers with psychiatric disabilities. WEIS items constitute a uni-dimensional construct given that the summary statistics for both item and person had a MNSQ of 1.00 and 1.02 respectively and both $\underline{t}$ values were -0.2. However, three items exceeded the established criterion for being too informative given the MNSQ < 0.7 and a $\underline{t}<-2.0$., suggesting these needed to be revised to enhance the quality measurement of the instrument. One hundred percent of the workers fit the expected response pattern of the Rasch model suggesting that workers with greater satisfaction, performance and health had a higher degree of match with his/her occupational environment. In summary, anecdotal data suggested that the WEIS provided clinically relevant information useful for planning of work-related interventions or reasonable accommodations.

Genetic characterization of pathogenic Leptospira species by DNA hybridization.

A total of 66 serovars of potentially pathogenic Leptospira species were examined by slot blot hybridization, and 57 of these serovars were classified in six DNA homology groups. In cases in which common serovars were studied, the results were in general agreement with the results of previous workers, who used different DNA homology methods. However, we propose a new species, Leptospira kirschneri, comprising the following serovars: bulgarica, butembo, cynopteri, dania, grippotyphosa, kabura, kambale, ramisi, and tsaratsovo. Seven of these serovars have not had their DNAs studied by other workers.

Alkylation damage, DNA repair and mutagenesis in human cells.

17 human cell lines that differ significantly in level of O6-alkylguanine-DNA alkyltransferase (AGT) activity were identified by comparing their sensitivity to the cytotoxic effect of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and determining the level of AGT activity in cell extracts from the various lines by measuring the decrease in radiolabeled O6-methylguanine from DNA, using high-performance liquid chromatography. 9 lines exhibited high levels of AGT activity, 2 showed an intermediate level (25-50% of the mean of those with the higher levels), and 6 exhibited very low or virtually undetectable levels of AGT. Included were several lines that are very deficient in capacity for nucleotide excision repair. When representatives from the 3 categories of cell lines defined by the level of AGT activity were compared for sensitivity to the cytotoxic and mutagenic effect of MNNG, they showed an inverse correlation between the degree of cell killing and frequency of mutants induced and the level of AGT activity. The cells' capacity for nucleotide excision repair did not affect these results. Exposure of cells with a high level of AGT activity to O6-methylguanine in the medium reduced the AGT activity 60-80%. These pre-treated cells exhibited a significantly higher frequency of MNNG-induced mutants than did cells that were not pre-treated, suggesting that the O6-methylguanine lesion in DNA is responsible for a significant proportion of the mutations induced. Cell strains containing substrates for assaying intrachromosomal homologous recombination were constructed using parental cell lines from each of the 3 categories of AGT activity. These strains showed an inverse correlation between the level of AGT activity and the frequency of MNNG-induced recombination. When various cell lines representing the 3 categories of AGT activity were compared for sensitivity to ethylnitrosourea, the results were consistent with AGT and nucleotide excision repair playing a role in preventing cell killing and mutation induction by this agent.

DNA relatedness among strains of Leptospira biflexa.

The slot blot method of DNA hybridization was used to study 38 strains of Leptospira biflexa belonging to 38 serovars. Fifteen of these serovars were placed into six groups. The remaining 23 serovars were generally too diverse to show significant DNA relatedness either to these groups or to one another. Serovar thracia was related to Group 5, but it was not included in this group because its percent relatedness was too low. We found that genetically related organisms were antigenically dissimilar. The absence of any significant genetic relationship between Leptonema illini and the Leptospira biflexa serovars tested supports the placement of the former species in a separate genus.

Fibroblasts from patients with hereditary cutaneous malignant melanoma are abnormally sensitive to the mutagenic effect of simulated sunlight and 4-nitroquinoline 1-oxide.

Because of a possible etiologic link between mutations and carcinogenesis, we compared fibroblasts derived from skin biopsies of several patients with hereditary cutaneous malignant melanoma and the dysplastic nevus syndrome for sensitivity to the mutagenic and/or cytotoxic effect of broad-spectrum simulated sunlight and of a UV mimetic carcinogen, 4-nitroquinoline 1-oxide (4NQO). The genetic marker was resistance to 6-thioguanine; loss of colony-forming ability was the assay for cytotoxicity. All five strains tested were more sensitive than normal to the killing effect of 4NQO (slopes of survival curves were 2- to 3-fold steeper), but only one strain was hypersensitive to killing by Sun Lamp radiation. Two strains were tested for mutagenicity. The response of each to the mutagenic action of these agents corresponded to its response to cell killing. Both strains were hypermutable after exposure to 4NQO, but only one showed a higher than normal frequency of mutants induced by simulated sunlight. The finding that nonmalignant fibroblasts from patients with a hereditary variant of malignant melanoma are abnormally susceptible to carcinogen-induced mutations suggests that hypersensitivity to mutagens contributes to risk of melanoma in patients. It also supports the somatic cell mutation hypothesis for the origin of cancer.

An in situ assay for induced diphtheria-toxin-resistant mutants of diploid human fibroblasts.

The sensitivity of diploid human fibroblasts to the cytotoxic effects of diphtheria toxin (DT) depended on cell growth status. Exponentially growing cells treated with 10(-3)-1 lethal flocculating units (LF) of DT/ml for 4 days survived with a frequency of 4 x 10(-4). However, the DT-resistant phenotype of colonies isolated under these conditions was not stable. When the growth of the cells had been arrested by confluence or deprivation of serum growth factors prior to treatment with DT (4 days, 10(-3)-0.6 LF/ml), the survival decreased to 2 x 10(-6) and the resistance of isolated colonies was stable. An in situ assay for induced DT-resistant mutants was developed in order to avoid problems associated with the possible reduced viability of the mutants relative to that of wild-type cells. A reproducible and linear dose response was obtained for the induction of DT-resistant mutants by ethylnitrosourea. The mutants were induced with high frequency by this compound (e.g., 10(-3) mutants/viable cell at a 37% survival dose); complete expression of the mutant phenotype occurred after 6 generations of growth under nonselective conditions. Isolated mutant colonies showed stable resistance to DT and were cross-resistant to Pseudomonas aeruginosa exotoxin A.