Nutrient and faecal contamination and retention in wetland enclosures (gei wais) in the Mai Po Marshes,Hong Kong

Nutrient and faecal contamination is an increasing problem to the shrimp productivity and wildlife conservation at the internationally important wetland ecosystems of the Mai Po Marshes (Hong Kong, P.R. China). The present study examined the nutrient status and faecal bacteria loading and potential retention capacity of contaminants of two wetland enclosures. Water in the wetland enclosures was eutrophicated with high concentrations of dissolved inorganic nitrogen (inorg–N_diss= 15.0 mg l^–1) and orthophosphate phosphorus (o-P = 1.89 mg l^–1) and was loaded with high levels of faecal coliforms (172 ×10³ cfu in 100 ml) and faecal streptococci (1.94 ×10³ cfu in 100 ml). The pattern of nutrient enrichment of two wetland enclosures is related to a north-to-south pollution gradient from the Shenzhen River to the wetlands. By retaining tidal water in the wetland for an 8-day period, water quality was greatly improved; NH₄–N was removed by 83%, o-P by 45% and faecal bacteria by 100%. This implies a self-purification capability of the wetland enclosures and a potentiality of using them as an alternative sewage treatment.     

Continuous feeding of antimicrobial growth promoters to commercial swine during the growing/finishing phase does not modify faecal community erythromycin resistance or community structure

Aims: To investigate the effect of continuous feeding of antimicrobial growth promoters (tylosin or virginiamycin) on the swine faecal community. Methods and Results: The study consisted of two separate on‐farm feeding trials. Swine were fed rations containing tylosin (44 or 88 mg kg^?1 of feed) or virginiamycin (11 or 22 mg kg^?1 of feed) continuously over the growing/finishing phases. The temporal impact of continuous antimicrobial feeding on the faecal community was assessed and compared to nondosed control animals through anaerobic cultivation, the analysis of community 16S rRNA gene libraries and faecal volatile fatty acid content. Feeding either antimicrobial had no detectable effect on the faecal community. Conclusions: Erythromycin methylase genes encoding resistance to the macrolide–lincosamide–streptogramin B (MLS_B) antimicrobials are present at a high level within the faecal community of intensively raised swine. Continuous antimicrobial feeding over the entire growing/finishing phase had no effect on community erm‐methylase gene copy numbers or faecal community structure. Significance and Impact of the Study: Antimicrobial growth promoters are believed to function by altering gut bacterial communities. However, widespread MLS_B resistance within the faecal community of intensively raised swine likely negates any potential effects that these antimicrobials might have on altering the faecal community. These findings suggest that if AGP‐mediated alterations to gut communities are an important mechanism for growth promotion, it is unlikely that these would be associated with the colonic community.     

Development of a polyclonal antibody-based AC-ELISA and its comparison with PCR for diagnosis of canine parvovirus infection

A polyclonal antibody-based antigen-capture ELISA (AC-ELISA) has been developed for detection of Canine parvovirus (CPV) antigens in faecal samples of dogs. The assay uses rabbit anti-CPV polyclonal antibody as the capture antibody, guinea pig anti-CPV polyclonal antibody as tracing antibody and anti-guinea pig HRPO conjugate as the detection system. The optimum dilution of the capture antibody and the tracing antibody capable of detecting the CPV-2 antigens was found to be 1:1 600 and 1:400, respectively, in the check-board titration. In this study, a total of 152 samples (129 faecal samples and 23 cell culture supernatant) were tested both by AC-ELISA and by polymerase chain reaction (PCR). Of the samples tested, 69 and 78 samples were found positive by AC-ELISA and PCR, respectively. The AC-ELISA had relative sensitivity, relative specificity and accuracy of 88.4%, 100.0% and 91.4% respectively. The analytical sensitivity of AC-ELISA was estimated to be 10^2.8 TCID₅₀/mL whereas PCR sensitivity was 10^0.8 TCID₅₀/mL. The AC-ELISA is a simple, quick and reliable method for screening large numbers of faecal samples of dogs suspected of CPV infection.     

Evaluation of titanium dioxide and chromic oxide as digestibility markers in ponies fed alfalfa hay in relation to marker dosing frequency

In equines, Cr₂O₃ is widely accepted as an indigestible marker, but there are health concerns regarding the carcinogenic properties of Cr₂O₃. Recently, TiO₂ has been suggested to be an alternative digestibility marker in equines. However, a comparison between Cr₂O₃ and TiO₂ has not been made in equines. Six Welsh pony geldings (initial BW: 254±3 kg; 7 years of age) fed chopped alfalfa hay were used to evaluate the use of TiO₂ (Ti) and Cr₂O₃ (Cr) as markers for calculating apparent digestibility and to investigate the effect of frequency of marker administration on the measurement of digestibility values. Diets contained 4.65 kg dry matter (DM) chopped alfalfa hay supplemented with minerals, vitamins, TiO₂ (3.3 g Ti/day) and Cr₂O₃ (3.2 g Cr/day). Ponies were dosed with either 3.3 g Ti and 3.2 g Cr once daily (DF1) or with 1.65 g Ti and 1.60 g Cr twice daily (DF2). After adaptation to the diets and procedures for 14 days, voluntary voided faeces were collected quantitatively over 7 days and analysed for moisture, ash, Ti and Cr. Apparent total tract DM digestibility (DMD) and organic matter digestibility (OMD) were calculated using the total faecal collection (TFC) and marker method (Ti and Cr). The overall mean cumulative faecal recovery of Cr and Ti (as % of intake) were 102.0% and 96.6%, respectively. Mean daily faecal recoveries of Cr as well as of Ti were not different (P=0.323; P=0.808, respectively) between treatments. Overall daily faecal recovery of Cr differed (P=0.019) from 100% when the marker was dosed once daily, whereas overall daily faecal recovery was similar to 100% for both administration frequencies when Ti was used as a marker. For both markers, the coefficient of variation of the mean faecal marker recovery between horses was lower when the markers were administrated twice per day. Across treatments, cumulative DMD and OMD estimated with Ti were similar (P=0.345; P=0.418, respectively) compared with those values determined by TFC method. When Cr was used, the calculated cumulative DMD tended (P=0.097) to be greater compared with those estimated with TFC, and cumulative OMD values were overestimated (P=0.013). Orally supplemented Ti recovery in the faeces of ponies fed chopped alfalfa hay with Ti administered once or twice daily was close to 100%, making it the preferred marker for digestibility trials in equines.     

Gas Production by Intestinal Microflora of Lactose Maldigesters

To investigate whether the gas produced by faecal contents of lactose maldigesters is traceable to symptoms of intolerance we analysed by gas chromatography faecal extracts of frozen stool samples for their gas composition. The samples were taken from 21 patients with biopsy-proven hypolactasia, 10 with severe and 11 with mild symptoms (cumulative score), and were incubated in three different carbohydrate broths. After incubation, H₂, CO₂, air and methane contents were analysed in triplicate. Differences between patient groups with mild and severe symptom scores were found, but they did not reach statistical significance. Substantial intragroup variations in gas production among patients with severe (range of H₂in galactose: 0.9–8.3%, and range of CO₂: 5.9–15.9%) as well as with mild symptoms (H₂: 0–8.3%, and CO₂1.4–14.5%) were recorded. Two patients with severe and one with mild symptoms produced detectable amounts of methane. The individual symptoms, i.e. flatulence or abdominal pain, did not directly correlate with gas production, but abundant gas production had a tendency to associate with a greater difference in waistline measurements before and after lactose provocation. Preliminary comparisons between faecal H₂and CO₂production and bacterial groups isolated from the samples showed that high concentrations of Escherichia coli tended to be more often associated with high gas production. Numbers of lactobacilli and H₂concentrations showed a trend towards negative correlation.     

Comparisons of Sampling Procedures and Time of Sampling for the Detection of Salmonella in Danish Infected Chicken Flocks Raised in Floor Systems

Bacteriological follow-up samples were taken from 41 chicken (Gallus gallus) flocks in floor systems, where Salmonella enterica (Salmonella) had been detected either directly in bacteriological samples or indirectly by serological samples. Three types of follow-up samples were compared to each other within each flock: 1) 5 pairs of socks, analysed as 5 samples, 2) 2 pairs of socks, analysed as one sample, and 3) 60 faecal samples, analysed as one pooled sample. Agreement between sampling methods was evaluated by the following statistical tests: 'Kappa', 'The adjusted rand', McNemar's test for marginal symmetry, Proportion of agreement P₀, P₊, P_-, and Odds Ratio. The highest agreement was found between the 2 types of sock sampling, while the lowest agreement was found by comparing 60 faecal samples with 5 pairs of socks. Two pairs of socks analysed as one pool appeared to be just as effective in detecting S. enterica as the 60 faecal samples. In broiler flocks, 5 pairs of socks were used both in the routine samples taken at about 3 weeks of age for the establishment of infection of the flock, and as one of the follow-up samples taken shortly before slaughter age, which means that the only notable differences between the 2 sampling rounds were the age of the broilers and of their litter. S. enterica was detected more frequently in samples from broilers about 3 weeks old, than in similar samples taken from broilers a few days prior to slaughter at ca. 33–40 days of age.     

The roles of enteric bacterial sialidase,sialateO-acetyl esterase and glycosulfatase in the degradation of human colonic mucin

Sialidase activity in normal faecal extracts showed a preference for mucin-related glycoprotein and oligosaccharide substrates, but the presence of two or moreO-acetyl esters at positions C₇–C₉ on the sialic acids retarded the rate of hydrolysis. A specific sialateO-acetyl esterase was detected with a lower total activity relative to sialidase with mucin substrates and having a pH optimum of 7.8 and aK _M of approximately 1mm sialateO-acetyl ester. A specific glycosulfatase activity was found in faecal extracts using the substrate lactit-[³H]ol 6-O-sulfate with a pH optimum of pH 5.0 and aK _M of approximately 1mm.Faecal extracts from ulcerative colitis (UC) patients had higher sialateO-acetyl esterase and glycosulfatase activity, while mucin sialidase activity was unchanged.Metabolically labelled mucin isolated from UC patients contained less sulfate and had lower sialic acidO-acetylation compared with normal mucin. Colonic mucin was degraded more efficiently by faecal extracts from UC patients compared with normal extracts. The UC mucin was degraded more rapidly than the normal mucin by faecal enzyme extracts from both normal and UC subjects. Abbreviations: UC, ulcerative colitis; BSM, bovine submandibular gland mucin; PMSF, phenylmethylsulfonyl-fluoride. Sialic acids are abbreviated according to Schauer [37].     

Isolation of bacteriophage host strains of Bacteroides species suitable for tracking sources of animal faecal pollution in water

Microbial source tracking (MST) methods allow the identification of specific faecal sources. The aim is to detect the sources of faecal pollution in a water body to allow targeted, efficient and cost‐effective remediation efforts in the catchment. Bacteriophages infecting selected host strains of Bacteroides species are used as markers to track faecal contaminants in water. By using a suitable Bacteroides host from a given faecal origin, it is possible to specifically detect bacteriophages of this faecal origin. It can thus be used to detect specific phages of Bacteroides for MST. With this objective, we isolated several Bacteroides strains from pig, cow and poultry faeces by applying a previously optimized methodology used to isolate the host strains from humans. The isolated strains belonged to Bacteroides fragilis and Bacteroides thetaiotaomicron. These strains, like most Bacteroides species, detected phages of the Siphoviridae morphology. Using the newly isolated host strains for phage enumeration in a range of samples, we showed that these detect phages in faecal sources that coincide with their own origin (70–100% of the samples), and show no detection or very low percentages of detection of phages from other animal origins (from 0 to 20% of the samples). Only strains isolated from pig wastewater detected phages in 50% of human sewage samples. Nevertheless, those strains detecting phages from faecal origins other than their own detected fewer phages (2–3 log₁₀ pfu·100 ml^?1) than the phages detected by the specific strain of the same origin. On the basis of our results, we propose that faecal source tracking with phages infecting specific Bacteroides host strains is a useful method for MST. In addition, the method presented here is feasible in laboratories equipped with only basic microbiological equipment, it is more rapid and cost‐effective than other procedures and it does not require highly qualified staff.     

Reliability of δ13C and δ15N in faeces for reconstructing savanna herbivore diet

We tested the reliability of herbivore faecal δ¹³C and δ¹⁵N values for reconstructing diet through review of an extensive database derived from a 3-year study of ungulates in South Africa's Kruger National Park. Faeces are a useful material for stable isotope studies of diet because they record dietary turnover at very short time scales, and because sampling is non-invasive. However, the validity of faecal isotope proxies may be questioned because they represent only undigested food remains. Results from Kruger Park confirm that free-ranging browsers have faecal δ¹³C consistent with C₃ feeding, grazer faeces are C₄, and mixed-feeder faeces intermediate. Although the respective ranges do not overlap, there is significant variation in faecal δ¹³C of browsers and grazers (～2.0–4.0‰) across space and through time. We demonstrate that most (～70%) of this variation can be ascribed to corresponding patterns of variation in the δ¹³C of C₃ and C₄ plants, respectively, re-enforcing the fidelity of faecal isotope proxies for diet but highlighting a need for mixing models that control for variations in plant δ¹³C in order to achieve accurate diet reconstructions. Predictions for the effects of climate (rainfall) and ecophysiology on ¹⁵N-abundance variations in mammals do not persist in faeces. Rather, faecal δ¹⁵N tracks changes in plant δ¹⁵N, with further fractionation occurring primarily due to variations in dietary protein (reflected by %N). Controlling for these effects, we show that a dual-isotope multiple source mixing model (Isosource) can extend diet reconstructions for African savanna herbivores beyond simplified C₃/C₄ distinctions, although further understanding of variations in mammal δ¹⁵N are needed for greater confidence in this approach.     

A dual marker technique to estimate individual feed intake in young pigs

Accurate estimation of individual feed intake (FI) of pigs could help better understand the variation in performance between individual animals. We studied dual marker methods to estimate individual FI in pigs. This method is based on the measurement of the ratio between two indigestible markers in faeces. Twelve 6.5-week-old individually housed male pigs were assigned to one of three oral dosing treatments supplying 180 mg of ytterbium chloride (YbCl₃)/day and 111 mg of dotriacontane (C32)/day as reference markers, either once (R1), three times (R3) or five times (R5) daily. Pigs were offered a diet containing 0.46 g/kg of chromium chloride (CrCl₃) and 0.15 g/kg of hexatriacontane (C36) as in-feed markers. The experiment lasted for 10 days: days ?5 to 0: adaptation; days 1–3: dosing of reference marker; days 2–4: total faecal collection. Spot faecal samples were taken on day 3 at 1200 h, 1700 h and on day 4 at 0700 h. Pigs were fed restrictedly three times daily, at 133.6 g/kg BW^0.60. Individual measured FI was recorded daily and was compared to predicted FI using the ratio of the dual marker pairs (Yb:Cr and C32:C36), both in total faecal collection and spot samples. Due to unequal variance, R1 pigs were omitted from the statistical treatment comparison. When using total faecal collection samples, the absolute prediction error (APE) (predicted FI minus measured FI) in R3 and R5 pigs was numerically lower than in R1 pigs, regardless of the marker pair used. The APE measured by C32:C36 was numerically lower than measured by Yb:Cr at all frequencies, and significantly (P = 0.039) in R3 pigs (C32:C36: 0.15 ± 0.02 kg/day; Yb:Cr: 0.29 ± 0.04 kg/day). This was related to a larger difference in faecal recovery between Yb and Cr compared with C32 and C36. Daily total faecal collection revealed that for R3 pigs, starting faecal collections 2 days after the onset of provision of the reference marker improved the APE when compared with starting after 1 day. When using C32:C36 to predict feed intake, pooled, but not single spot samples gave similar APEs compared with total faecal collections. Therefore, we recommend dosing the reference marker three times per day for 2 days on days 1 and 2, combined with pooled spot faecal sampling collected on days 3 and 4. In this way, absolute prediction errors of 10%-15% of simultaneously measured intakes of multiple nutrient resources in a complex housing system are feasible using the dual marker technique.     

Inactivation of virus during anaerobic digestion of manure in laboratory scale biogas reactors

Reduction of porcine parvovirus, bovine enterovirus and faecal enterococci were measured in biogas reactors continuously run on manure and manure supplemented with household waste at 35°C and 55°C and in batch test run at 70°C. The aim of the experiments was to study the sanitation effect of anaerobic digestion and to evaluate the use of faecal enterococci as an indicator of sanitation. Parallel studies on the reduction of virus and faecal enterococci were done in physiological saline solution. Heat ínactivation was found to be an important factor in thermophilic biogas plants and the overall dominant factor at 70°C. However, other environmental factors with a substantial virucidal and bactericidal effect were involved in inactivation. The death rates for faecal enterococci were generally higher than for porcine parvovirus and lower than for bovine enterovirus. For faecal enterococci, a logarithmic reduction of 4 (corresponding to the recommended minimum guaranteed retention time) was obtained after 300 hours at 35°C and after 1–2 hours at 55°C. In continuously-fed reactors, a high reduction rate was initially seen for the virus tested, followed by a reduction in the rate. For porcine parvovirus, a minimum guaranteed retention time of 11–12 hours is necessary at 55°C in the initial phase (0–4 hours) and 54 hours hereafter (4–48 h). Correspondingly, for bovine enterovirus a MGRT of 23 hours was necessary at 35°C and < 0.5 hours at 55°C. The data indicate that faecal enterococci measurements give a good indication of inactivation of enterovirus and other more heat sensitive virus, especially under thermophilic conditions. Parvovirus is very suitable for comparative investigations on inactivation in the temperature range of 50–80°C, due to the extreme thermal resistance of this virus. However, in stipulating sanitation demands for biogas reactors it seems more reasonable to use less resistant virus, such as a reovirus or picornavirus, which better represents the pathogenic animal virus.Abbreviations BEV bovine enterovirus - CFU colony forming unit - FE faecal enterococci - HRT hydraulic retention time - MGRT minimum guaranteed retention time - ND not detected - PPV porcine parvovirus - TCID₅₀ tissue cell infective dose 50 % - VFA volatile fatty acids - VS volatile solids     

Radiocaesium variability within sheep flocks: determination of 137Cs intake in free-ranging sheep

A study is described in which ¹³⁷Cs intake by free-ranging sheep was estimated at two farms in the area of west Cumbria (northwest England) which received some of the highest amounts of fallout from the Chernobyl accident within the United Kingdom. The faecal excretion of ¹³⁷Cs was estimated from faecal ¹³⁷Cs activity concentrations and the use of intraruminal controlled release devices containing Cr₂O₃ to determine faecal dry matter output. The intake of ¹³⁷Cs was estimated by assuming an apparent absorption coefficient appropriate to the herbage grazed. The methodology has the advantage that sampling of herbage representative of that ingested by study animals is not required. Caesium-137 dietary intake explained >60% of the observed variability in the ¹³⁷Cs activity concentration determined in the muscle of sheep. Resultant transfer coefficient (F _f ) values to describe the transfer of ¹³⁷Cs from the diet to muscle were in agreement with previously reported values. At one farm, there was a positive correlation between the ¹³⁷Cs activity concentration in muscle and F _f whilst at the other farm there was a negative correlation between F _f and ¹³⁷Cs dietary intake. Potential reasons for these observations are discussed. Received: 6 April 2000 / Accepted: 25 May 2000     

Plasma cortisol and faecal cortisol metabolites concentrations in stereotypic and non-stereotypic horses: do stereotypic horses cope better with poor environmental conditions?

Background

Stereotypic behaviours, i.e. repetitive behaviours induced by frustration, repeated attempts to cope and/or brain dysfunction, are intriguing as they occur in a variety of domestic and captive species without any clear adaptive function. Among the different hypotheses, the coping hypothesis predicts that stereotypic behaviours provide a way for animals in unfavourable environmental conditions to adjust. As such, they are expected to have a lower physiological stress level (glucocorticoids) than non-stereotypic animals. Attempts to link stereotypic behaviours with glucocorticoids however have yielded contradictory results. Here we investigated correlates of oral and motor stereotypic behaviours and glucocorticoid levels in two large samples of domestic horses (N _Study1 = 55, N _Study2 = 58), kept in sub-optimal conditions (e.g. confinement, social isolation), and already known to experience poor welfare states. Each horse was observed in its box using focal sampling (study 1) and instantaneous scan sampling (study 2). Plasma samples (collected in study 1) but also non-invasive faecal samples (collected in both studies) were retrieved in order to assess cortisol levels.

Results

Results showed that 1) plasma cortisol and faecal cortisol metabolites concentrations did not differ between horses displaying stereotypic behaviours and non-stereotypic horses and 2) both oral and motor stereotypic behaviour levels did not predict plasma cortisol or faecal cortisol metabolites concentrations.

Conclusions

Cortisol measures, collected in two large samples of horses using both plasma sampling as well as faecal sampling (the latter method minimizing bias due to a non-invasive sampling procedure), therefore do not indicate that stereotypic horses cope better, at least in terms of adrenocortical activity.     

Application of long-chain alcohols as faecal markers to estimate diet composition of horses and cattle fed with herbaceous and woody species

Utilization of long-chain alcohols (LCOH) as diet composition markers in horses and cattle was assessed in a study conducted with 12 mature crossbreed mares (385±47 kg BW) and six adult non-lactating cows (499±36 kg BW) of Asturiana de los Valles breed. The LCOH data were combined with alkane and long-chain fatty acid (LCFA) data to test the applicability of combining these markers to estimate diet composition. Animals were randomly divided into groups of three animals and received a daily total amount of 1.0 kg dry matter/100 kg BW of diets composed of different proportions of ryegrass (Lolium perenne) and woody species (Ulex gallii and heather). Diet composition was estimated from even-chain LCOH (C₂₀-OH to C₃₀-OH) combined or not with alkane (C₂₅-C₃₁ and C₃₃) and/or LCFA (C₂₂-FA to C₂₈-FA, C₃₀-FA, C₃₂-FA and C₃₄-FA) concentrations in diet components and faeces by least-squares procedures, using marker faecal concentrations uncorrected for incomplete faecal recovery (FR0) or corrected using mean recoveries across diets within animal species (FR1). Results showed large differences between plant species in their LCOH profiles, and that these markers offered additional discriminatory information to that provided by alkanes and LCFA. The LCOH markers were incompletely recovered in the faeces of both animal species. In cattle, LCOH FR tended to increase with carbon-chain length in a linear manner in both diets (P<0.001), whereas in horses overall data showed a curvilinear relationship between these variables. Combination of LCOH, LCFA and alkanes resulted in more accurate diet estimates. Correction of faecal LCOH concentrations to incomplete FR led to more accurate diet composition estimates in both animal species. Results obtained in this study suggest the usefulness of LCOH markers combined with alkanes and LCFA to estimate diet composition of horses and cattle grazing mixed grassy–woody plant communities.     

Measurements of digestibilities of cholesterol and fatty acids using 5α-cholestane as an inert marker in the tilapia, Oreochromis niloticus, and the freshwater prawn, Macrobrachium rosenbergii

An attempt was made to measure the apparent digestibility (AD) of cholesterol and fatty acids using 5aL-cholestane as a marker (cholestane-method) using tilapia. Oreochromis niloticus, and the freshwater prawn, Macrobrachium rosenbergii, as test animals. The results were compared with those obtained by the Cr₂O₃-method. Casein-based test diets containing 0.3% levels of cholestane and chromic oxide as markers were prepared for determination of AD of dietary cholesterol and fatty acids. After 2 weeks of acclimation, the test animals, tilapia (average 3.0 g wet weight) and freshwater prawn (average 0.5 g wet weight) were fed the test diets for 1 week, and faecal samples were collected over 6 subsequent days. Lipids in the diets and faeces were extracted with chloroform-methanol and saponified with 10% KOH in methanol. Gas-liquid chromatography (GLC) on 1.5% OV-17 of the unsaponifiable matters afforded the quantities of 5aL-cholestane and cholesterol, whereas that on 5% Shinchrom E-71 of the saponifiable matters provided the quantities of fatty acids. Chromic oxide was determined by a wet-digestion method. In tilapia, the AD of cholesterol determined by cholestane- and Cr₂O₃-methods were 78.1% and 73.5%, respectively. The AD of individual fatty acids differed with the types of fatty acids, but those of fatty acids determined by the two methods in this study were similar, except for a slightly higher value of several fatty acids such as 16:1 in the cholestane-method than in the Cr₂O₃-method. This indicates that 5α-cholestane as well as Cr₂O₃ can be conventionally used as a marker for determining AD of fatty acids and cholesterol. The cholestane-method is advantageous as it permits parallel analysis of cholesterol (or fatty acids) and 5α-cholestane by GLC using small amounts of faecal sample. The cholestane-method was also useful for analysing digestibilities of cholesterol and total lipids in freshwater prawn. However, slight differences between the cholestane- and Cr₂O₃-methods were observed in the AD of several fatty acids such as 14:0 and 16:1.     

A 4 year observation of gastrointestinal nematode egg counts,nemabiomes and the benzimidazole resistance genotypes of Teladorsagia circumcincta on a Scottish sheep farm

Anthelmintic resistance threatens the sustainability of sheep production globally. Advice regarding strategies to reduce the development of anthelmintic resistance incorporates the outcomes of modelling exercises. Further understanding of gastrointestinal nematode species diversity, and population dynamics and genetics (which may vary between species) is required to refine these models; and field studies combining faecal egg outputs, species composition and resistance genetics are needed to calibrate them. In this study, faecal samples were taken from ewes and lambs on a commercial farm in south-eastern Scotland at approximately 3 t-4 week intervals between spring and autumn over a period of 4 years. Faecal egg counts were performed on these samples, and L₃ were collected from pooled coprocultures. Deep amplicon sequencing was used to determine both the species composition of these L₃ and the proportions of benzimidazole-resistant single nucleotide polymorphisms in the isotype-1 β-tubulin locus of the predominant species, Teladorsagia circumcincta L₃. Despite consistent management throughout the study, the results show variation in gastrointestinal nematode species composition with time and between age groups, that was potentially associated with weather conditions. The F200Y benzimidazole resistance mutation is close to genetic fixation in the T. circumcincta population on this farm. There was no evidence of variation in isotype-1 β-tubulin single nucleotide polymorphisms frequency between age groups, and no genetic evidence of reversion to benzimidazole susceptibility, despite targeted benzimidazole usage. This study highlights the need to include speciation when investigating gastrointestinal nematode epidemiology and anthelmintic resistance, and serves as an example of how genetic data may be analysed alongside species diversity and faecal egg counts, when markers for other anthelmintic classes are identified.     

Abundance and germination capability of resting cysts of Alexandrium spp. (Dinophyceae) from faecal pellets of macrobenthic organisms

The Alexandrium spp. resting cysts were found abundantly in faecal pellets collected from the bottom sediments at two stations in Hiroshima Bay. It is considered that these faecal pellets were excreted by the macrobenthos, such as polychaeta and mollusca, based on their size and morphology. Polychaeta was the most dominant macrobenthos, and mollusca was the second most dominant group in Hiroshima Bay. The resting cysts of Alexandrium spp. in the bottom sediments at the two stations were counted in both the faecal pellets of macrobenthos and in the surrounding sediment. As a result, the number of cysts in the faecal pellets accounted for 28.9-35.2% of total cysts. In addition, cysts isolated from faecal pellets had almost the same germination ability as those in the sediment. Thus, Alexandrium cysts are tolerant to the predation and digestive processes of macrobenthic organisms.     

Quantitative PCR assays to survey the bovine adenovirus levels in environmental samples

Aims: Previous studies suggested bovine adenoviruses (BAdVs) could be used as cattle faecal indicators. The main aim of this study was to survey the levels of BAdV in environmental samples using quantitative polymerase chain reaction (qPCR). Methods and Results: Two qPCR assays were developed to identify and quantify BAdVs in environmental samples. BAdVs were detected in all dairy manure, and in most cases, the sample concentrations were around 10³–10⁴ copies ml^?1. Farm tile drainage samples were also detected, but the concentrations were about 1–3 log₁₀ lower than the BAdV concentrations in the manure samples. The genome equivalent copy (GEC) levels of BAdV and the phage forming unit (PFU) levels of somatic phage in manure samples were comparable. Four of twenty individual cattle faeces were positive with concentrations similar to that found in the manure samples. Sequencing results confirmed the presence of BAdV in the environmental samples, and phylogenetic analysis indicated that BAdV 2 and 4 were the most prevalent serotypes in all the manure samples tested. The qPCR assays developed in this study showed higher sensitivity in detecting BAdV 1 and 2 than the previous published nested assay. Conclusion: The high levels of BAdV in the environmental samples may suggest it could be used for bovine faecal indicator. The significant levels of BAdV in the drainage samples may indicate the potential of surface water pollution by the manure applied to farm fields. Significance and Impact of the Study: This is the first study that reports the quantitative level of BAdV in environmental samples. These results could be useful when it comes to determining whether BAdV could be utilized as a bovine faecal indicator.     

Antemortem detection of mouse parvovirus and mice minute virus by polymerase chain reaction (PCR) of faecal samples

Parvoviruses remain one of the most common viral infections seen in laboratory mouse colonies. The purpose of this study was to develop an antemortem polymerase chain reaction (PCR) assay to detect mice infected with mouse parvovirus-1 (MPV) and mice minute virus (MMV) using faecal samples. The MMV PCR assay consistently detected as few as 100 plasmid copies of MMV in faecal samples, while the MPV PCR assay detected as few as 10 plasmid copies of MPV. Faecal pellets from infected mice held at room temperature from 1 to 7 days tested positive by MMV and MPV PCR, respectively. This demonstrates that parvovirus DNA is stable in faecal samples kept at room temperature. PCR assays were also used to follow the length of MMV and MPV shedding in faeces from SENCAR mice, which were endemically infected with multiple agents. MMV faecal shedding was detected in 60-70% of the mice 5-7 weeks old, and by 13 weeks of age, faecal samples from all mice were negative for MMV. MPV faecal shedding was detected in 90-100% of the mice 5-11 weeks old; however, by 19 weeks of age, faecal samples from all mice were negative for MPV. These findings confirm that faecal shedding occurs for a limited time and suggest that 5-9-week-old mice are the most appropriate age group in endemically infected mice for faecal testing by MMV and MPV PCR.     

Gastrointestinal and immunological responses of senior dogs to chicory and mannan-oligosaccharides

Thirty-four senior dogs (pointers 8 - 11 years, beagles 9 - 11 years) were used to evaluate the effects of oligosaccharides on nutritional and immunological characteristics. Dogs were randomly allotted to treatments [1% chicory (CH), 1% mannan-oligosaccharide (MOS), 1% chicory + 1% MOS (CM), or no supplementation (control, CON)] in a parallel design with a 4 week baseline period followed by a 4 week treatment period. Dietary supplementation with MOS or CM tended (P = 0.07) to increase food intake due, in part, to an increase in fermentable fibre and a decrease in energy content of the diet. Although wet faecal output increased (P < 0.05) for dogs supplemented with MOS or CM, when corrected for food intake, no differences were noted. The CM treatment increased (P < 0.05) faecal score (1 = hard and dry, 5 = watery liquid), although these scores remained in a desirable range (3 to 3.5). Chicory supplementation increased (P = 0.07) fat digestibility. Chicory or MOS increased (P  0.05) faecal bifidobacteria concentrations 0.4 and 0.5 log₁₀ cfu/g DM, respectively, compared to the CON, while MOS decreased (P < 0.05) faecal E. coli concentrations. Oligosaccharides did not affect white blood cell (WBC) concentrations, but CH and CM tended to increase (P = 0.10) neutrophil concentrations compared to control dogs. Peripheral lymphocyte concentrations were decreased in dogs supplemented with MOS (P = 0.06) and CM (P < 0.05). Chicory and MOS alter faecal microbial populations and certain indices of the immune system of senior dogs.