Fecal Pats Research Articles

Modelling E. coli runoff concentrations from sheep and cow grazed pastures in New Zealand: challenges and future research needs

ABSTRACT A model of Escherichia coli concentrations in the runoff from grazed pastures was developed to increase our knowledge and ability to predict the relative impact of both cow and sheep grazing on water quality. Input parameters to the initial model were derived from published data and model outputs were compared with measured E. coli runoff concentration data from both sheep and cow grazed pasture. The model was designed to represent the complexities of year-round rotational grazing farm system. For cow grazing, the model estimated runoff E. coli concentrations in the same order of magnitude as the measured data. However, for sheep grazing, the model underestimated the measured runoff E. coli concentrations by 1.5 orders of magnitude. To understand this negative result, Monte Carlo simulation techniques were used to conduct a sensitivity analysis of the input parameters. This analysis identified that runoff concentrations were highly sensitive to inputs of the E. coli concentrations in the dung and the mobilisation rates of E. coli from the dung. This modelling study demonstrates that significant gaps and uncertainties remain in our understanding and prediction of the mobilisation and transport of E. coli from grazed pasture systems. HIGHLIGHTS Faecal microbial losses from agriculture are poorly understood. A runoff concentration model was developed based on the area of faecal pats. The area-based model appears to work for cow grazed pastures. The area-based model does not work for sheep grazed pastures.

Comparison of Three Preharvest Sampling Strategies to Monitor Pathogens in Cattle Lairage Areas

The objective of this study was to compare preharvest monitoring strategies by evaluating three different sampling methods in the lairage area to determine pathogen recovery for each sampling method and incoming pathogen prevalence from the cattle to inform in-plant decision making. Samples were gathered over a 5-month period, from February to June 2022, at a harvesting and processing facility located in Eastern Nebraska. Sampling methods included (i) fecal pats, (ii) boot swabs, and (iii) MicroTally swab. A total of 329 samples were collected over the study period (fecal pats: n = 105, boot swabs: n = 104, and MicroTally swabs: n = 120). Specific media combinations, an incubation temperature of 42°C, and incubation timepoints (18–24 h) were utilized for each matrix and the prevalence of Salmonella, Escherichia coli O157:H7, and six non-O157 Shiga-toxin producing E. coli (STEC) was evaluated using the BAX system Real-Time PCR assay. Overall, results from the study concluded that boot swabs were an effective sampling method for pathogen detection in the cattle lairage area. Boot swabs (97.1%) were statistically more likely to detect for Salmonella (p < 0.05) when compared to fecal pats (67.6%) and MicroTally swab (77.5%) methods. For E. coli O157:H7 and STEC – O26, O121, O45, and O103 prevalence, boot swabs were significantly better at detecting for these pathogens (p < 0.05) than MicroTally swabs (OR = 3.16 – 11.95) and a comparable sampling method to fecal pats (OR = 0.93 – 2.01, p > 0.05). Lastly, all three sampling methods detected a very low prevalence for E. coli O111 and O145; therefore, no further analysis was conducted. The boot swab sampling method was strongly favored because they require little training to implement, are inexpensive, and they do not require much sampling labor; therefore, would be a simple and effective sampling method to implement within the industry to evaluate pathogen prevalence preharvest.

Effect of Different Anthelmintic Drugs on the Development and Efficacy of Duddingtonia flagrans

Nematophagous fungi are a biological control tool used against gastrointestinal nematodes in livestock. These fungi prey on free-living larvae in faeces and could be affected by active drugs excreted post-treatment. This study aimed to determine in vitro and under environmental conditions the effect of the following anthelmintics on the fungus Duddingtonia flagrans: ivermectin, levamisole, albendazole, fenbendazole and ricobendazole. The in vitro effect of anthelmintics on fungal growth and predatory capacity was assessed in corn meal agar and coprocultures, respectively. Ivermectin (1, 2 and 10 ppm), fenbendazole (0.027, 0.054 and 1 ppm) and albendazole (1 ppm) significantly affected fungal development. The fungal efficacy against L3 was high in the control and levamisole coprocultures but decreased significantly in the presence of albendazole, fenbendazole, ricobendazole and ivermectin. The impact of levamisole on D. flagrans was further assessed under environmental conditions in autumn and winter; the fungal efficacy measured in faecal pats and the surrounding herbage was not affected by levamisole at any time. This study shows that using albendazole, fenbendazole, ricobendazole or ivermectin may compromise fungal activity, as these drugs affect the free-living stages of nematodes in faeces, but levamisole can be safely considered in parasite control strategies involving D. flagrans and anthelmintic treatments.

PSVI-27 Evaluation of Coated and Non-Coated Seed Species Pertinent to the Great Basin in Response to in Vitro Ruminal Incubation

Abstract Target grazing has exhibited itself as an effective method of invasive weed mitigation across degraded rangelands in the Great Basin. Additionally, if the seeds of target species are supplemented into the diets of ruminants, they can serve as vectors for the dispersal of native seed species through fecal pats, which may prove to be a useful tactic for rangeland rehabilitation. However, a challenge associated with seed supplementation, is that the seeds of many target species are sufficiently degraded in the rumen. Therefore, the aim of this study was to evaluate the effects of seed coating technologies on germination rates post ruminal in vitro incubation. In vitro incubations were performed on seven species of seeds pertinent to the Great Basin: Achnatherum hymenoides (ACHY), Achillea millefolium (ACMI), Agropyron cristatum (AGCR), Atriplex canascens (ATCA), Elymus elymoides (ELEL), Pseudoregneria spicata (PSSP), and Sphaeralcea grossulariifolia (SPRT) uncoated (control) or coated with either Polyvinylpyrrolidone (PVP) or PVP+ ethocel, as well as uncoated Bromus tectorum (BRTE) seeds. A Daisy II incubator (Ankom Technology, Macedon, NY) was used for the in vitro analysis with rumen inoculum from two rumen-cannulated crossbred Angus steers (body weight = 910.5 ± 34.5 kg) and a buffer ratio of 1:4. Two F57 filter bags containing 1 g of seeds per treatment (control, PVP or PVP+ethocel) were then placed inside of the Daisy II incubator and kept at 39°C for 12 and 24 h for the control treatments, and for 12, 24, and 48 h for the PVP and PVP+ethocel coating treatments. For uncoated seeds, ACHY germination by 24 h was greater when compared with 0 h (P &amp;lt; 0.05), whereas ATCA germination rate decreased linearly by 12 h (P &amp;lt; 0.05). No differences were observed over time on germination rates of BRTE, as well as all of the other uncoated seeds (P &amp;gt; 0.05). When seeds were coated with PVP, ATCA germination rates started to linearly decrease by 12 h (P &amp;lt; 0.05), whereas PVP coated ACMI germination rates only started to decrease by 24 h (P &amp;lt; 0.05). The germination rates of PVP coated ELEL and PSSP seeds only started to significantly decrease by 48 h (P &amp;lt; 0.05). When coated with PVP+ethocel, the observed germination rates were similar to those observed for PVP coated seeds. However, when compared with uncoated seeds, coating with PVP+ethocel seem to decrease germination rates of ACMI and ATCA by 0 h (P &amp;lt; 0.05). No changes on germination rates were observed over time for coated and uncoated AGCR and SPRT (P &amp;gt; 0.05). In conclusion, seed germination rates after in vitro incubation were not increased by coating the seeds with PVP or PVP+ethocel.

353 Evaluation of Native and Invasive Seed Germination Ability After in Situ Incubation in the Rumen for Rangeland Restoration Through Targeted Seed Supplementation

Abstract Changes to native plant compositions of drylands are often associated to loss of wildlife, increased frequency, and severity of wildfires; further, it complicates the grazing permits and allocation during periods of restoration. The goals herein investigate the germination rates of native seed species after in situ incubation to evaluate their potential dispersal as supplements for grazing cattle. Native: Indian ricegrass (Achnatherum hymenoides; ACHY), white yarrow (Achillea millefolium; ACMI), crested wheatgrass (Agropyron cristatum; AGCR), sagebrush (Artemisia tridentata; ARTR), fourwing saltbrush (Atriplex canascens; ATCA), bottlebrush squirreltail (Elymus elimoides; ELEL), bluebunch wheatgrass (Pseudoregneria spicata; PSSP), and gooseberryleaf globemallow (Sphaeralcea grossulariifolia; SPRT) and nonnative: cheatgrass (Bromus tecorum, BRTE; fall BRTEf and spring seeds BRTEs) seeds were examined. Four ruminally canulated Angus x Hereford steers adapted to a forage kochia-alfalfa diet were utilized for seed incubations. Seeds were weighed into 84 bags at a rate of 5 g per bag (duplicated within and in different animals). Seeds were incubated sequentially (0, 6, 12, 24, 36, 48, and 96 h) and removed at the 96-h point when seeds were thoroughly washed and immediately counted (50) on square 80 mm 80 mm commercial sterile petri dishes with moisture pads and 5 mL of water. Seeds were incubated at 20 °C with 12 h of light/no-light intervals. Germination plates were incubated for 50 d and counted every 10 d for germinations. Generalized linear models assuming a Poisson distribution with separation of means through Dunnett’s multiple comparisons test. Statistical significance was declared at P &amp;lt; 0.05. All statistical analyses were performed on R (R Core Team 2022). Incubation times inside of the rumen linearly decreased germination rates across most seed species. For BRTEs, germination completely stopped at 48 h with a steep decrease observed after 24 h incubation. The BRTEf were also incubated but showed low germination both before and after rumen exposure (winter dormancy). Though ARTR is very important in rangeland restoration in the intermountain west, its germination declined more than 85% within just a 6 h incubation in situ (P &amp;lt; 0.001). A significant germination decrease was observed for AGCR at 36 h; however, germination was full at 48 h and 50% of the original at 96 h (P &amp;lt; 0.05). For ACMI, germination rates significantly declined after 36 h incubation, and for PSSP after 24 h. Low germinations were observed for ACHY, SPRT, and ATCA even with high reported in-bag tetrazolium chloride viabilities. Our results highlight that cattle grazing of cheatgrass does not spread seeds through fecal pats. Additional tests are required to determine seed viability of the dormant (low germination) BRTEf. Seed toleration to ruminal degradation highlights the potential targeted seed supplementation during grazing for rangeland restoration.

Antimicrobial resistance of Escherichia coli from dairy farms participating in an antimicrobial stewardship educational program for farm employees

Antimicrobial use in food-producing animals is under increasing scrutiny due to the potential effect on the selection of antimicrobial-resistant bacteria that may be transmitted to humans by direct contact, with the food chain, or the environment. Novel data monitoring commensal Escherichia coli from dairy farms is essential for understanding antimicrobial resistance (AMR) patterns and their association with herd health management practices. The objectives of this study were to: (1) compare the prevalence of antimicrobial resistance in the E. coli isolates from the hospital, fresh, and mid-lactation pens from 18 conventional dairy farms participating in an educational training program in antimicrobial stewardship practices in California and Ohio, and (2) to characterize the prevalence of antimicrobial resistance of commensal E. coli isolated from pooled fecal pat samples before and 3 mo after participating in the educational training program. Pooled fecal pat samples were collected from the hospital pen, the fresh pen (1 to 5 DIM), and the mid-lactation pens (90 to 150 DIM) on conventional dairies in CA (n = 9) and OH (n = 9). Fecal samples were collected as part of a larger study using a quasi-experimental design that assigned farms to the training intervention group (TG; 6 per state) or the control group (CG; 3 per state). For the TG, farm worker(s) identified as having the task of diagnosis and treatment of adult cows on the farm participated in a training program on antimicrobial stewardship practices. Pooled fecal samples (n = 7) were collected at enrollment and 3 mo after completing the intervention on each of the participating farms (n = 18), followed by culture for E. coli isolation and antimicrobial sensitivity testing using the broth microdilution methodology. Logistic regression models were used to evaluate the association between E. coli antimicrobial resistance patterns with the training intervention and farm-level factors. No effect was observed in the prevalence of resistant isolates between the control and intervention farms after the training was delivered. Isolates from the hospital pens were 2.48 (95% CI: 1.06-6.22, P = 0.03) and 5.61 (95% CI: 1.94-16.91, P < 0.001) times, more likely to be resistant to streptomycin and chloramphenicol, respectively, than isolates from the mid-lactation pens. Our findings indicate there was a higher prevalence of AMR in E. coli associated with the hospital pen within the farm, while the training program for 3 mo did not affect the prevalence of AMR in E. coli on the farms participating in the program. Further research efforts should be conducted to identify factors driving AMR at the pen level, as well as approaches that could be used to reduce the risk of disseminating AMR from sick pens to animals being housed and to other pens on the farm.

Biotic potential and pathogenicity of a Haemonchus contortus susceptible laboratory isolate compared to a resistant field isolate in Brazil

The high prevalence of anthelmintic resistance of Haemonchus contortus is threatening sheep farming. In this context, resistant populations’ replacement using susceptible isolates appears to be a strategy to overcome this situation. The aim of the present study was to evaluate the ability of a susceptible H. contortus laboratory isolate in comparison with a multiresistant field isolate to infect lambs and to study its behaviour in field conditions. Sixteen worm-free lambs were experimentally infected with 4000 infective larvae (L3) of a H. contortus anthelmintic-resistant field isolate (Group R, n = 8) or a laboratory isolate susceptible to anthelmintics (Group S, n = 8). Groups were placed in separate clean pastures, where they were kept for 266 days. Faecal egg counts (FEC) showed maximum values between 21- and 42-days post-infection (PI) and decreased over time, with no statistical differences between groups (P > 0.05). The pathogenicity of the isolates was similar, as lambs from both groups showed a decrease in packed cell volume and total plasma protein values in the beginning of the study, coinciding with the highest FECs. Production of L3 were similar for both groups, when evaluating faecal cultures kept in vitro or L3 recovered from pasture samples and from faecal pats collected from the paddocks, which suggests that both isolates have the same potential of development in vitro and in the environment. The degree of resistance/susceptibility to thiabendazole (TIA), levamisole (LEV), monepantel (MON) and ivermectin (IVE) was evaluated using the Larval Development Assay (RESISTA-Test©) 160 days PI. The resistant isolate showed a high resistance factor (RF) to TIA (141), LEV (822), MON (34) and IVE (196), while the RF of the susceptible isolate remained below three, which indicates high susceptibility to all drugs tested. Our study demonstrates that the susceptible isolate was able to infect lambs and did not show higher pathogenicity nor any differences on its biotic potential when compared to the resistant field isolate. These results indicate that the susceptible isolate has the potential to be used to replace a resistant population of H. contortus, which should be validated in long-term studies under commercial sheep farming conditions.

Trapping Behaviour of Duddingtonia flagrans against Gastrointestinal Nematodes of Cattle under Year-Round Grazing Conditions.

The purpose of using nematophagous fungi as biological control agents of gastrointestinal nematodes of livestock is to reduce the build-up of infective larvae on pasture and thus avoid clinical and subclinical disease. As the interaction of fungus-larval stages takes place in the environment, it is crucial to know how useful the fungal agents are throughout the seasons in areas where livestock graze all year-round. This study was designed to determine the predatory ability of the nematophagous fungus Duddingtonia flagrans against gastrointestinal nematodes of cattle during four experiments set up in different seasons. In each experiment, faeces containing eggs of gastrointestinal nematodes were mixed with 11,000 chlamydospores/g and deposited on pasture plots. A comparison between fungal-added faeces and control faeces without fungus were made with regard to pasture infectivity, larval presence in faecal pats, faecal cultures, faecal pat weight, and temperature inside the faecal mass. In three of the four experiments, Duddingtonia flagrans significantly reduced the population of infective larvae in cultures (68 to 97%), on herbage (80 to 100%), and inside the faecal pats (70 to 95%). The study demonstrated the possibility of counting on a biological control tool throughout most of the year in cattle regions with extensive grazing seasons.

Survival, mobilization, and transport of Escherichia coli from sheep faeces.

This study looked at the survival, mobilization, and transport of Escherichia coli from sheep faeces over an extended period, which was then repeated for all four seasons. Rain simulation was used to measure E. coli mobilization directly from faecal pats and subsequent transport across a soil surface. The rain simulation experiments were conducted over the same 90-day period and for the four seasons as per the survival experiments conducted to determine E. coli survival in faecal pats. The survival experiments demonstrated up to three orders of magnitude increase of E. coli in sheep faeces. Peak E. coli concentrations in the faeces were observed on Day 2 through to Day 30, with no significant effect of seasons or weather patterns. The E. coli concentrations in the runoff followed the survival pattern of E. coli concentrations in the faeces. There was a statistically significant relationship between the E. coli concentrations in the faeces and the runoff. The data generated on the relationship between Escherichia coli concentrations in the faeces and in the runoff can support the development of runoff risk models for sheep grazing pastureland to support catchment modelling, land use decisions, and public health.

Evaluation of Strategies to Reduce Equine Strongyle Infective Larvae on Pasture and Study of Larval Migration and Overwintering in a Nordic Climate.

Horses, as grazing animals, are inadvertently exposed to intestinal parasites that, if not controlled, may cause disease. However, the indiscriminate use of anthelmintic drugs has led to drug resistance, highlighting the need for pasture-management practices to reduce the level of parasitic exposure and lessen reliance on drugs. The efficacy of such methods depends both on the epidemiology of the parasites and the prevailing weather conditions. The aim of the study was to investigate the effect of faecal removal and harrowing on reducing the number of parasite larvae in herbage. Moreover, the migratory and survival ability of strongyle larvae in a Nordic climate was studied. Faeces from horses naturally infected with strongyle nematodes were used to contaminate pastures and grass samples were collected to harvest larvae. Twice-weekly faecal removal significantly reduced larval yields, whereas harrowing on a single occasion under dry weather conditions in the summer did not. Strongyle larvae were able to migrate 150 cm from the faecal pats, but most larvae were found within 50 cm. Both Cyathostominae and S. vulgaris survived the winter months with larvae harvested up to 17-18 months after faecal placement. Resting of pastures for one year greatly reduced the parasite level, but two years of rest were required for parasite-free pasture.

Effect of a direct-fed microbial (10-G Armor) on feedlot performance, carcass characteristics, and prevalence of Salmonella in fed-beef heifers.

Crossbred beef heifers (N = 1,394; initial shrunk body weight [BW] 291 ± 9.9 kg) were used to investigate the efficacy of 10-G Armor (Life Products, Inc., Norfolk, NE; 10-G) upon feedlot performance, carcass characteristics, and fecal and subiliac lymph nodes Salmonella prevalence. Heifers were blocked by day of arrival and allocated to 1 of 20 pens (N = 70 heifers/pen) and assigned one of two treatments (10 pens/treatment): no direct-fed microbial (CON) or 2 g/heifer/d of Lactobacillus acidophilus, Enterococcus faecium, Pediococcus pentosaceus, Lactobacillus brevis and Lactobacillus plantarum, respectively (Life Products, Inc., Norfolk, NE; 10-G). Twenty-four animals were randomly selected from each pen for Salmonella sampling. Recto-anal mucosal swab samples (RAMS) were obtained at initial processing and harvest; subiliac lymph nodes were collected at harvest. In addition, pen surface fecal pats were collected and composited by pen (10 pats per composite, 5 composites per pen) on days 0, 52, 120, and 192. Data were analyzed as a generalized complete block design, and pen served as the experimental unit. No differences were observed in live growth performance metrics (P ≥ 0.55). Yield grade distributions did not differ between treatments (P ≥ 0.62); however, cattle fed 10-G tended (P = 0.06; 14.6% vs. 18.9%) to have fewer USDA Select carcasses and more (P = 0.09; 73.6% vs. 78.0%) USDA Choice carcasses. Cattle fed 10-G tended (P = 0.10; 9.2% vs. 12.3%) to have fewer liver abscesses and had fewer (P = 0.04; 5.3% vs. 8.5%) severe liver abscesses. Salmonella prevalence of RAMS did not differ between treatments at initial processing (P = 0.97; CON = 11.6%, 10-G = 11.5%) or at harvest (P = 0.91; CON = 99.0%, 10-G = 98.6%); however, RAMS differed (P < 0.01) in Salmonella prevalence between the two collection times. Cattle fed 10-G had a lower frequency of Salmonella positive lymph nodes (P = 0.01; CON = 15.8%, 10-G = 7.4%) than CON. However, Salmonella log (mpn/g) of lymph nodes did not differ between treatments at harvest (P = 0.34; CON = 0.73, 10-G = 0.34). These data indicate that cattle fed 10-G have decreased rates of severe liver abscesses without altering live animal performance or carcass characteristics. Supplementation of 10-G significantly reduced the prevalence rate of Salmonella recovered from the subiliac lymph nodes. The factors responsible for the observed difference in the effects of 10-G on Salmonella warrant further investigation

Monacrosporium sinense and Pochonia chlamydosporia for the biological control of bovine infective larvae in Brachiaria brizantha pasture

Ecologically correct solutions are necessary to minimize the use of anthelmintics in the control of cattle parasitic diseases. This study evaluated the effects of oral administration pellets containing the fungal combination Monacrosporium sinense and Pochonia chlamydosporia in the biological control of infective larvae (L3) of parasitic gastrointestinal nematodes of cattle grazed in pasture. Twelve crossbred bovines (Holstein × Zebu), seven to nine years old, after dewormed, were randomly divided into two groups (group treated with the combination of fungi and control group) and placed in separate paddocks of Brachiaria brizantha that were naturally infested with L3. Feces samples were collected from the animals to determine the parasitic load and pasture samples were collected from the soil to determine the L3 infestation. The recovery of L3 from the pasture of the treated group was 91.7% and 86.4% less than in the pasture of the control group, respectively for samples collected at 20 cm and 40 cm from the faecal pats. The values of eggs per gram of feces in the months of May 2019 and January 2020 were lower (p ≤ 0.05) in the treated group with the fungal combination. The combined administration of M. sinense and P. chlamydosporia reduced the L3 infestation on pasture and in animal parasitic load, making it a tool for controlling bovine gastrointestinal parasitic nematodes.

Salmonella Prevalence Varies Over Time and Space in Three Large, Adjacent Cattle Operations in the Southwestern United States

We set out to describe the prevalence of Salmonella enterica in three large, adjacent cattle operations in the southern High Plains of the United States. Operations included two dairies (one of which routinely administers a commercially available Salmonella vaccine) and one feedlot. Samples were collected monthly for 12 months. At each sample collection, 25 freshly voided fecal pats and a sample from each of the water troughs were collected from each of five pens of cattle within an operation. Each monthly collection included a total of 375 fecal and ~32 water samples for a yearly total of 4,500 and 379 samples, respectively (note that the number of water troughs per pen varied within an operation). Salmonella was commonly recovered from fecal (71.3%) and water (28.5%) samples and tended to follow somewhat similar temporal patterns over time. However, its prevalence varied among operations despite being adjacent properties in that Salmonella was recovered from 61.3, 80.1, and 75% of fecal samples from dairy 1, dairy 2 and the feedlot, respectively. Salmonella prevalence in water samples across collection times averaged 36.1, 70.2, and 46.1% for dairy 1, dairy 2, and the feedlot, respectively. While it is uncertain why the Salmonella prevalence varied from operation to operation, the higher observed prevalence of Salmonella in water on dairy 2 and/or the use of a commercial Salmonella vaccine by dairy 1 may offer a partial explanation.

Factors influencing the detection of antibacterial-resistant Escherichia coli in faecal samples from individual cattle.

To investigate whether on-farm antibacterial usage (ABU), environmental antibacterial-resistant (ABR) Escherichia coli prevalence, sampling and sample handling methodologies are associated with ABR E. coli positivity in individual faecal samples from dairy heifers. Three hundred and sixty-four heifers from 37 farms were sampled via rectal or faecal pat sampling. Samples were stored at -80°C for variable periods before microbiological analysis. Data analysis was done through a multilevel, multivariable logistic regression approach. Individual rectal samples had increased odds of positivity for amoxicillin-, cefalexin- and tetracycline-resistant E. coli. Sample storage for 6-12months was associated with decreased odds of finding amoxicillin- and tetracycline-resistant E. coli. On-farm ABU had little influence, and environmental ABR E. coli prevalence had no significant influence on the odds of sample-level positivity for ABR E. coli. Sampling methodology and sample handling have a greater association than on-farm factors with the detection of ABR E. coli in individual faecal samples from dairy heifers. Sampling and storage methodologies should be considered carefully at the point of designing ABR surveillance studies in livestock and their environments and, where possible, these methodologies should be standardized between and within future studies.

Seasonal epidemiology of gastrointestinal nematodes of cattle in the northern continental climate zone of western Canada as revealed by internal transcribed spacer-2 ribosomal DNA nemabiome barcoding

Background Gastrointestinal nematode (GIN) epidemiology is changing in many regions of the world due to factors such as global warming and emerging anthelmintic resistance. However, the dynamics of these changes in northern continental climate zones are poorly understood due to a lack of empirical data.Methods We studied the accumulation on pasture of free-living infective third-stage larvae (L3) of different GIN species from fecal pats deposited by naturally infected grazing cattle. The field study was conducted on three organic farms in Alberta, western Canada. Grass samples adjacent to 24 fecal pats were collected from each of three different pastures on each farm. Internal transcribed spacer-2 nemabiome metabarcoding was used to determine the GIN species composition of the harvested larvae. The rotational grazing patterns of the cattle ensured that each pasture was contaminated only once by fecal pat deposition. This design allowed us to monitor the accumulation of L3 of specific GIN species on pastures under natural climatic conditions without the confounding effects of pasture recontamination or anthelmintic treatments.ResultsIn seven out of the nine pastures, grass L3 counts peaked approximately 9 weeks after fecal deposition and then gradually declined. However, a relatively large number of L3 remained in the fecal pats at the end of the grazing season. Nemabiome metabarcoding revealed that Cooperia oncophora and Ostertagia ostertagi were the two most abundant species on all of the pastures and that the dynamics of larval accumulation on grass were similar for both species. Daily precipitation and temperature across the whole sampling period were similar for most of the pastures, and multiple linear regression showed that accumulated rainfall 1 week prior to sample collection had a significant impact on the pasture L3 population, but accumulated rainfall 3 weeks prior to sample collection did not.Conclusions The results suggest that the pasture L3 population was altered by short-term microclimatic conditions conducive for horizontal migration onto grass. Overall, the results show the importance of the fecal pat as a refuge and reservoir for L3 of cattle GIN on western Canadian pastures, and provide an evidence base for the risk assessment of rotational grazing management in the region.Graphical

Assessment of Contamination Risk from Fecal Matter Presence on Fruit and Mulch in the tomato fields based on generic Escherichia coli population

The purpose of this study was to evaluate the fate of generic Escherichia coli in fecal pats under different field conditions and to predict Salmonella and Shiga toxin-producing E. coli (STEC) survival dynamics based on developed models. Eight trials were conducted during spring and fall in both North and Central Florida. Fresh cattle feces (1g) was placed on mature green round tomatoes located inside (TIP) and outside (TOP) of the plant canopy. Fecal pats (10 g) were placed under (MUP) and distant (MDP) to tomato plants on plastic mulch. Pathogen populations were predicted based on developed models. Declines in generic E. coli populations over 7 days (Log CFU/g) in fecal pats were between 0.9 and 2.7 on TIP, 1.2 and 3.0 on TOP, 0.2 and 1.2 on MUP, and 0.4 and 1.5 on MDP in the Central Florida fall and spring trials, respectively. E. coli populations remained stable at ≤4.2 and ≤ 6.5 Log CFU/g during all North Florida trials. The concentration changes in predicted Salmonella and STEC population were less than 2.1 Log CFU/g in fecal pats for all conditions. Developed models predicted similar pathogen survival trends to generic E. coli with no dramatical impact under the field conditions.

Environmental risk assessment of veterinary parasiticides used in cattle

All veterinary medicines have to undergo a rigorous environmental risk assessment before gaining a marketing authorisation; post-marketing pharmacovigilance ensures constant surveillance for any environmental impacts that had not been anticipated from earlier research. Products are labelled to reflect any potential risk points. Despite these procedures, some compounds, notably parasiticides of the macrocyclic lactone class that are used in large animal practice, have gained a reputation among some scientists, the general public and the media for having adverse effects on dung insect fauna and for causing delays in the degradation of faeces on pasture. While the toxicity of faecal residues of insecticides to non-target insect fauna, when measured under controlled conditions in short-term experiments is indisputable, the consequences of such activity at ecological scales are much less well defined. Dung provides an ephemeral habitat for an enormous array of macro- and micro-invertebrates, fungi and bacteria, whose composition changes as pats age and disintegrate. Perturbations to such complex systems inevitably have a variety of outcomes and this is reflected in field trials, which can produce conflicting results. This article summarises pertinent aspects of the underlying biology and ecology of the bovine faecal pat and the evidence for significant impact of veterinary medication.

High levels of third-stage larvae (L3) overwinter survival for multiple cattle gastrointestinal nematode species on western Canadian pastures as revealed by ITS2 rDNA metabarcoding

BackgroundThe ability of infective larvae of cattle gastrointestinal nematode (GIN) species to overwinter on pastures in northerly climatic zones with very cold dry winters is poorly understood. This is an important knowledge gap with critical implications for parasite risk assessment and control.MethodsInfective third-stage larvae (L3) were quantified in samples of fecal pats, together with adjacent grass and soil, before and after winter on three farms in southern, central and northern Alberta. Nemabiome ITS2 metabarcoding was then performed on the harvested L3 populations to determine the species composition. Finally, parasite-free tracer calves were used to investigate if the L3 surviving the winter could infect calves and develop to adult worms in spring.ResultsFarm level monitoring, using solar powered weather stations, revealed that ground temperatures were consistently higher, and less variable, than the air temperatures; minimum winter air and ground temperatures were − 32.5 °C and − 24.7 °C respectively. In spite of the extremely low minimum temperatures reached, L3 were recovered from fecal pats and grass before and after winter with only a 38% and 61% overall reduction over the winter, respectively. Nemabiome ITS2 metabarcoding assay revealed that the proportion of L3 surviving the winter was high for both Cooperia oncophora and Ostertagia ostertagi although survival of the former species was statistically significantly higher than the latter. Nematodirus helvetinaus and Trichostrongylus axei could be detected after winter whereas Haemonchus placei L3 could not overwinter at all. Adult C. oncophora, O. ostertagi and N. helvetianus could be recovered from tracer calves grazing after the winter.ConclusionsThe largest proportion of L3 were recovered from fecal pats suggesting this is important refuge for L3 survival. Results also show that L3 of several GIN parasite species can survive relatively efficiently on pastures even in the extreme winter conditions in western Canada. Tracer calf experiments confirmed that overwintered L3 of both C. oncophora and O. ostertagi were capable of establishing a patent infection in the following spring. These results have important implications for the epidemiology, risk of production impact and the design of effective control strategies. The work also illustrates the value of applying ITS2 nemabiome metabarcoding to environmental samples.

Environmental sampling to assess the bioburden of Mycobacterium avium subspecies paratuberculosis in drylot pens on California dairies.

Mycobacterium avium subspecies paratuberculosis (MAP) is a bacterium that can cause substantial economic losses in infected dairy herds due to reduced milk production and increased cow-replacement costs. In order to control MAP in dairies with drylot pens, a standardized environmental sampling protocol to quantify MAP in fecal slurry was developed based on an existing protocol for freestall pens. Specifically, following a 24 h hold of the flush, a grab sample of approximately 10 ml of fecal slurry was collected every 1 m along the flush lane of the drylot pens, avoiding individual cow fecal pats. To determine the reliability and repatability of the new environmental sampling protocol for estimation of MAP bioburden at the pen level, two collectors simultaneously collected fecal slurry samples every day for 3 days from six drylot cow pens on two Central California dairies. During the study period no cow movement between pens was allowed with the exception of sick cows. The study herds had MAP seroprevalence of 5.8% and 3.2%, respectively, based on whole pen serum ELISA results. Variance components models for quantitative real-time PCR (qPCR) results showed samples collected from different pens on different dairies accounted for greater variablitiy in MAP concentration (65%), while samples collected by different collectors had the least variability (0.1%). In contrast, variability in MAP concentration in environmental samples collected on different days had 25% variability. The intraclass correlation coefficient showed high reliability (93%) of environmental sampling simultaneously by different collectors. In contrast, the reliability of environmental sampling at different days was 65%, which was similar to the reliability for sampling by different collectors on different days. Investigators can expect high reliability when employing the new environmental sampling protocol along with qPCR testing of environmental samples from drylot pens.

Stimulating Duddingtonia flagrans chlamydospore production through dehydration

Duddingtonia flagrans is a nematode-trapping fungus that has shown promising results as a tool to combat parasitic nematode infections in livestock. The fungus interrupts the parasitic lifecycle by trapping and killing larval stages on pasture to prevent re-infection of animals. One barrier to the fungus’ commercial use is scaling up production of the fungus, and specifically of chlamydospores, which survive the digestive tract to grow in fecal pats on pasture, thus have potential as a feed through anthelmintic. The purpose of this study was to evaluate the effect of dehydration on sporulation of the fungus. Disks of Duddingtonia flagrans type strain (ATCC® 13423™) were grown on 17% cornmeal agar for 26 days at 30 °C, then split into three groups; dried quickly at 38 °C and 37% humidity over 48 h (“incubated”), dried more slowly at 24 °C and 55% humidity over 10 days (“air-dried”), or kept at 30 °C and sealed with parafilm to prevent loss of moisture as a control (“wet”). Half of each dried culture was resuspended in water, then heated to liquify and homogenized through vortexing. Spores were then counted in a Neubauer hematocytometer. Both the “air-dried” and “incubated” drying techniques yielded significantly more spores than the “wet” control (Welch’s two sample t test p values of .0359 and .0411, respectively). The difference in average chlamydospores per milliliter was insignificant between the two drying techniques, although a visual representation of the data shows less spore count variability in the “air-dried” technique.