Concentration Of Protozoa Research Articles

Multi-omics in vitro study of the salivary modulation of the goat rumen microbiome.

Ruminants are able to produce large quantities of saliva which enter into the rumen and salivary components exert different physiological functions. Although previous research has indicated that salivary immunoglobulins can partially modulate the rumen microbial activity, the role of the salivary components other than ions on the rumen microbial ecosystem has not been thoroughly investigated in ruminants. To investigate this modulatory activity, a total of 16 semi-continuous in vitro cultures with oats hay and concentrate were used to incubate rumen fluid from four donor goats with autoclaved saliva (AUT) as negative control, saliva from the same rumen fluid donor (OWN) as positive control, and either goat (GOAT) or sheep (SHEEP) saliva as experimental interventions. Fermentation was monitored throughout 7days of incubation and the microbiome and metabolome were analysed at the end of this incubation by Next-Generation sequencing and liquid chromatography coupled with mass spectrometry, respectively. Characterisation of the proteome and metabolome of the different salivas used for the incubation showed a high inter-animal variability in terms of metabolites and proteins, including immunoglobulins. Incubation with AUT saliva promoted lower fermentative activity in terms of gas production (-9.4%) and highly divergent prokaryotic community in comparison with other treatments (OWN, GOAT and SHEEP) suggesting a modulatory effect derived from the presence of bioactive salivary components. Microbial alpha-diversity at amplicon sequence variant (ASV) level was unaffected by treatment. However, some differences were found in the microbial communities across treatments, which were mostly caused by a greater abundance of Proteobacteria and Rikenellacea in the AUT treatment and lower of Prevotellaceae. These bacteria, which are key in the rumen metabolism, had greater abundances in GOAT and SHEEP treatments. Incubation with GOAT saliva led to a lower protozoal concentration and propionate molar proportion indicating a capacity to modulate the rumen microbial ecosystem. The metabolomics analysis showed that the AUT samples were clustered apart from the rest indicating different metabolic pathways were promoted in this treatment. These results suggest that specific salivary components contribute to host-associated role in selecting the rumen commensal microbiota and its activity. These findings could open the possibility of developing new strategies to modulate the saliva composition as a way to manipulate the rumen function and activity.

In vitro aflatoxins recovery after changing buffer or protozoa concentrations in the rumen fermentation fluid.

This study simulates in vitro the effects of (i) rumen acidity and (ii) change in rumen protozoa numbers on the recovery of aflatoxins (AFs). Two 24-hfermentation experiments were carried out using the same batch in vitro fermentation systems and substrate (dried corn meal) containing 11.42, 2.42, 7.65and 1.70 µg/kg of AFB1, AFB2, AFG1and AFG2respectively. In Experiment 1,two buffer concentrations (normal salts dosage or lowered to 25%) were tested. Buffer reduction decreased gas production (730 vs. 1101 mL, p < 0.05), volatile fatty acids (VFA) and NH3 concentrations in the fermentation liquid (39.8 vs. 46.3 mmol/L, and 31.7 vs. 46.5 mg/dLrespectively,p < 0.01). Recovery of all four AFs types was higher (p < 0.01) in the reduced buffer fermentation fluid, both as a percentage of total AF incubated (73.6% vs. 62.5%, 45.9% vs. 38.1%, 33.6% vs. 17.9% and 18.9% vs. 6.24% for AFB1, AFB2, AFG1and AFG2respectively) and as amounts relative to VFA production (163.4 vs. 123.5, 22.1 vs. 15.7, 48.8 vs. 22.5 and 6.16 vs. 1.86 ng/100 mmol of VFA, for AFB1, AFB2, AFG1and AFG2respectively). In Experiment 2,Stevia rebaudiana Bertoni extracts (S) or a Camphor essential oil (Cam) were added to fermenters and compared to the control (no additives, C). S and Cam addition resulted in a 25% reduction (p < 0.05) and a 15% increase (p < 0.05) in protozoa countsrespectively, when compared to C. Both plant additives slightly reduced (p < 0.05) AFB1 recovery as a percentage of total AFB1 incubated (68.5% and 67.7% vs. 74.9% for S, Camand Crespectively).Recoveries of all other AFs were unaffected by the additives. In conclusion, the rumen in vitro AFB1 recovery (63%-75%) was higher than other AFs (3%-46%) and the acidic fermentation environment increased it. In our conditions, changes in protozoa numbers did not affect AFs recovery.

Inactivation of Giardia intestinalis cysts in water using a sonochemical method based on mid-high-frequency waves

Giardia intestinalis is a pollutant of food and water, resistant to conventional disinfection treatments and its elimination requires effective methods action. Herein, mid-high-frequency ultrasound (375 kHz), which produces HO• and H2O2, was used as an alternative method of treatment to inactivate Giardia intestinalis cysts in water. The effect of ultrasound power (4.0, 11.2, 24.4 W) on the sonogeneration of radicals was tested, showing that 24.4 W was the condition most favorable to treat the parasite. The viability of the protozoan cysts was evaluated using the immunofluorescence technique and vital stains, showing this protocol was useful to quantify the parasite. The sonochemical method (at 375 kHz and 24.4 W) was applied at different treatment times (10, 20, and 40 min). A significant decrease in the protozoan concentration (reduction of 52.4% of viable cysts) was observed after 20 min of treatment. However, the extension of treatment time up to 40 min did not increase the inactivation. Disinfecting action was associated with attacks on the Giardia intestinalis cyst by sonogenerated HO• and H2O2 (which may induce structural damage, even the cell lysis). For future work is recommended to test combinations with UVC or Fenton process to enhance the inactivating action of this method.•Mid-high-frequency ultrasound produces HO• and H2O2 profitable to inactivate Giardia intestinalis.•Immunofluorescence technique and vital stains allowed us to quantify the parasite viability.•Giardia intestinalis cysts concentration decreased by 52.4% after only 20 min of sonication.

Effects of rosemary (Rosmarinus officinalis) by-products and linseed (Linumusitatissimum) intake on digestibility, body weight gain, and estimated tissular composition in cull fat-tailed ewes.

The aim of this study was to evaluate the effect of rosemary (Rosmarinus officinalis) residues (RR) as dietary supplement in association with linseed (Linumusitatissimum) on digestive aspects and body weight (BW) gain in fat-tailed Barbarine ewes. To achieve this, 24 cull ewes (BW: 33 ± 2.7kg) were divided into four groups receiving a basal diet composed of 500g of straw for two groups (S) and 300g of straw in addition to 200g of RR for two other groups (R). All animals received 750g of concentrate; the usual concentrate (UC) was offered for one S (S-UC) and one R group (R-UC); the linseed concentrate (LC) was also offered for one S (S-LC) and one R group (R-LC). The fattening and digestibility trials lasted 90days. The dry matter (DM) intake and digestibility were similar across the groups. However, the crude protein (CP) intake and digestibility and the neutral detergent fiber intake were significantly affected by the treatments (P > 0.05). The nitrogen intake (NI), the fecal, and retained nitrogen were significantly affected by the regimen (P < 0.05); however, the urinary nitrogen was unchanged. The nitrogen balance was positive for all groups averaging 60% of the NI. The ruminal pH, ammonia nitrogen, and protozoa concentration were not affected by the RR intake and linseed supply; however, the control time affected significantly these parameters. Final BW, the average daily gain, the muscle, and fat contents were neither affected by the basal diet nor by the concentrate type (P > 0.05). In conclusion, the RR intake in association with linseed did not alter the animal's performances by improving their productive characteristics and consequently, the commercial value of this sheep's category.

Development and evaluation of a molecular based protocol for detection and quantification of Cryptosporidium spp. in wastewater

Infections caused by protozoan parasites are a major public health concern globally. These infections are commonly diagnosed during water-borne outbreaks, necessitating accurate and highly sensitive detection procedures to assure public health protection. Current molecular techniques are challenged by several factors, such as low parasite concentration, inefficient DNA extraction methods, and inhibitors in environmental samples. This study focused on the development and validation of a molecular protocol for DNA extraction, efficient protozoan (oo)cyst recovery and quantification of protozoan parasites from wastewater using droplet digital polymerase chain reaction (ddPCR). Five DNA extraction methods, including commercial kits, custom phenol-chloroform, and in-house modified methods, were evaluated. The efficiency of each method was assessed via spectrophotometric analysis and ddPCR amplification using specific primers. Lastly, the developed protocol was evaluated for the detection and quantification of Cryptosporidium parvum in wastewater from different regions in South Africa. The conventional phenol-chloroform extraction method yielded the highest DNA concentration of 223 (±0.71) ng/μl and detected the highest number of Cryptosporidium parvum (1807 (±0.30) copies/ddPCR reaction) compared to other methods evaluated in this study. Additionally, the phenol-chloroform method demonstrated high sensitivity in extracting DNA from as few as one cyst/L of Cryptosporidium parvum, corresponding to 5.93 copies/ddPCR reaction. It was also observed that analysis of both the filtered supernatant and pellets after centrifugation improves the recovery efficiency of oocysts from wastewater by 10.5%, resulting in a total recovery of 64.1%. This optimized protocol was successfully applied to measure protozoan concentration in wastewater from different regions in South Africa. The improved DNA extraction and quantification method proposed in this study would be effective in monitoring protozoan concentration in the environment, which will help in instituting mitigation measures to reduce water-borne infections.

Comparison of the microbial communities of alpacas and sheep fed diets with three different ratios of corn stalk to concentrate.

The objective of this study was to investigate the characteristics of ruminal microbial communities of alpacas (Lama pacos) and sheep (Ovis aries) fed three diets with varying ratios of roughage (corn stalk) to concentrate, 3:7 (LS), 5:5 (MS) and 7:3 (HS). Six alpacas (one-year-old and weighing 29.5±7.1kg) and six sheep (one-year-old and weighing 27.9±2.7kg) were used in this study, in a replicated 3×3 Latin square experiment. Total protozoa concentration was determined under the microscope; total fungi and methanogens were assessed using quantitative polymerase chain reaction and expressed as a percentage of total bacterial 16S rRNA gene copies; bacterial communities were investigated by targeted 16S rRNA gene (V3-V4 region) sequencing. The percentage of fungi was significantly higher in alpacas than in sheep under the LS diet, while the concentration of protozoa was significantly lower in alpacas under HS, MS and LS diets. The alpha diversity including Shannon, Chao l and ACE indices of bacterial communities was higher in alpacas than in sheep, under the LS diet. A total of 299 genera belonging to 22 phyla were observed in the forestomach of alpaca and sheep, with Bacteroidetes and Firmicutes dominating both animal species. Phyla Armatimonadetes and Fusobacteria, as well as 64 genera, were detected only in alpacas, whereas phyla Acidobacteria and Nitrospira, as well as 44 genera, were found only in sheep. The abundance of cellulolytic bacteria, including Butyrivibrio and Pseudobutyrivibrio, was higher in alpacas than in sheep under all three diets. These differences in the forestomach microbial communities partly explained why alpacas displayed a higher poor-quality roughage digestibility, and a lower methane production. Results also revealed that the adverse effects of high-concentrate diets (70%) were lesser in alpacas than in sheep.

Effect of Sulfur and Urea Fortification of Fresh Cassava Root in Fermented Total Mixed Ration on the Improvement Milk Quality of Tropical Lactating Cows.

The aim of the present research was to determine the influence of sulfur and urea combined with fresh cassava root in fermented total mixed ration (FTMR) on digestibility, fermentation in the rumen, blood metabolite, milk yield, and milk quality in tropical lactating dairy cows. Four mid-lactation Thai Holstein–Friesian crossbred cows were studied. Pre-experiment milk yield was 12.7 ± 0.30 kg/day, and the body weight was 495 ± 40.0 kg. Animals were evaluated in a 2 × 2 factorial in a 4 × 4 Latin square design to receive diets followed by: factor A, which was a dose of sulfur inclusion at 1.0% and 2.0%, and factor B, which was level of urea inclusion at 1.25% and 2.5% DM in FTMR. The hydrogen cyanide (HCN) concentrations reduced 99.3% to 99.4% compared with fresh cassava root when FTMR was supplemented with 1.0% and 2.0% sulfur, respectively. Intake of crude protein was increased based on urea level addition (p < 0.05). Blood thiocyanate concentration was increased by 21.6% when sulfur was supplemented at 2.0% compared to 1.0% (p < 0.05). There was no difference in protozoal concentration, whereas bacterial populations at 4 h after feeding were significantly greater by 6.1% with the FTMR supplemented with 2.0% sulfur and 2.5% urea (p < 0.01). Allantoin concentrations, excretion, absorption, and microbial crude protein showed significant interactions between sulfur levels and urea levels in cows fed diets supplemented with 2.0% sulfur and 2.5% urea (p < 0.05). The molar ratios of the volatile fatty acid (VFA) profile were affected by dietary FTMR (p < 0.01). Furthermore, propionic acid increased by 4.6% when diets were supplemented by 2.5% sulfur (p < 0.01). Milk fat and total solids increased when feed was supplemented with 2.0% sulfur and 2.5% urea (p < 0.05). The diets supplemented with 2.0% sulfur levels resulted in greater concentrations of milk thiocyanate (p < 0.05). The somatic cell count was significantly reduced throughout the experiment with increasing sulfur supplementation (p < 0.05). Animals fed diets supplemented with 2.0% sulfur exhibited a decreased somatic cell count by 18.3% compared with those fed diets supplemented with 1.0% sulfur. Thus, inclusion of 2.0% sulfur with 2.5% urea in FTMR containing fresh cassava root improved digestibility, ruminal fermentation, microbial crude protein synthesis, and milk qualities in dairy cows.

Effects of pomegranate peel extract on ruminal and post-ruminal in vitro degradation of rumen inoculum of the dairy cow

This experiment was carried out to study the effect of water extracted pomegranate peel extract (PE) on ruminal protein degradation and post-ruminal digestion in the dairy cow. PE was added at six levels of total phenolics (g/kg of the basal diet); 3.75 (PE1); 4.4 (PE2); 5.05 (PE3); 5.70 (PE4); and 6.35 (PE5). Rumen degradable crude protein (rdCP) decreased with PE addition (L < 0.0001), but total CP degradability (tdCP) was not affected. Compared to PE0, PE2, and PE3 diets showed higher (L = 0.054, Q = 0.029) digestibility of bypass CP (dBCP). Increasing levels of PE resulted in a decrease in proteolytic bacteria numbers (p < 0.0001). At PE4 and PE5 levels, total VFA and acetate concentrations linearly decreased compared to PE0. PE inclusion lowered the acetate:propionate ratio (L = 0.0001) and Ammonia-N production after 24 h (L = 0.0008) of incubation. The total number of protozoa, genera Dasytricha and Isotricha, and subfamilies Entodiniinae, Diplodiniinae, and Ophrioscolecinae decreased with increasing dietary PE concentration (p < 0.0001). The results suggest that all levels of PE addition reduce the protozoal population and Ammonia-N concentration. All PE levels slowed down protein degradation in the rumen but PE2 and PE3 showed the greatest effect.

Rumen protozoal dynamics during the transition from milk/grass to high-concentrate based diet in beef calves as affected by the addition of tannins or medium-chain fatty acids

Changes in rumen protozoal community during adaptation of 7-month-old beef calves from a milk/grass diet to a high-energy diet consisting of cereal-based concentrate plus wheat straw, both given ad libitum (diet C), were studied. Eighteen rumen-cannulated Limousine crossbred male calves were randomly assigned to three diets (C; C plus 20 g/kg of a 65:35 chestnut and quebracho tannin extract, T; and C plus 6 g/kg medium-chain fatty acid (MCFA) mixture, M). Fermentation variables were studied, and rumen ciliates were quantified and classified from rumen fluid sampled just before the morning feeding on days 0, 7, 14, 21 and 28 of the experiment. Protozoal population changed over time, but all calves harbored a stable population at the end of the experiment. Diversity decreased on time, and Entodinium and Isotricha species were the most abundant at the end of the trial. When occurred, defaunation was transient as protozoa were absent from M calves in three occasions on days 7 and 14 but were refaunated thereafter reaching a consistent population. Regardless the diet, proportions of Isotricha spp. and Dasytricha ruminantium, Epidinium spp. and Subfamily Diplodiniinae, decreased throughout the experiment, whereas Entodinium spp. increased reaching 0.93 of total protozoa. Dasytricha and protozoa from Subfamily Diplodiniinae disappeared from the rumen of all calves at the end of the study, except for Polyplastron multivesiculatum. Rumen variables were not affected by the addition of additives (P > 0.05). Rumen pH decreased across the study (P < 0.001) but did not reach acidotic values despite the high fermentability of the diet. Higher levels of lactate were observed on C calves at d14, but not in other dates (diet by day interaction, P = 0.03). Addition of tannin extract did not affect protozoal concentration and diversity, but MCFA might even lead to a transient defaunation. Transition to a high-concentrate diet does not necessarily reduce initial protozoal concentration, but diversity is affected towards a major proportion of Entodinium and, to a lesser extent, Isotricha. Dietary shift from weaning to a high-concentrate ration promotes a less diverse rumen protozoal population, but a high concentration is still maintained thereafter.

Soil warming and nitrogen deposition alter soil respiration, microbial community structure and organic carbon composition in a coniferous forest on eastern Tibetan Plateau

Understanding the response of soil respiration (Rs) to future climate projections is critical for understanding feedbacks and developing effective mitigation strategies. We conducted a warming and N deposition manipulation experiment using a 2 × 2 full-factorial design to evaluate the interactive effects of soil warming and N deposition on Rs, microbial community composition and soil organic carbon (SOC) composition in a subalpine coniferous forest on the eastern slope of the Qinghai-Tibetan Plateau. Soil warming and N addition independently increased Rs and changed microbial community composition, resulting in a net increase in bacteria with N addition and an increase in protozoa with warming; but the interaction of these two factors proved to be antagonistic, resulting in a lower Rs and protozoa concentration in the combined warming and nitrogen addition (WN) treatment relative to the warming (W) treatment. The chemical structure of SOC changed with the treatments, with alkyl C increased with warming, aromatic C decreased with both warming and N addition, and the interaction of warming and N induced a lower O-alkyl C and a higher contribution of alkyl C to SOC. The negative correlation between Rs and aromatic C and the positive correlation between Rs and alkyl C and MBC indicates that the elevated CO2 emission under warming and N addition was related to the decomposition of aromatic C and the following relative accumulation of alkyl C. Our observations suggest that climate induced soil warming will likely increase soil CO2 emissions, but this effect could be largely offset by a simultaneous increase in N deposition in subalpine coniferous forests.

Effects of Supplementation of Piper sarmentosum Leaf Powder on Feed Efficiency, Rumen Ecology and Rumen Protozoal Concentration in Thai Native Beef Cattle.

Simple SummaryThis work was to study the influence of Piper sarmentosum (PS) leaf powder supplementation in cattle diet. It was found that supplementation of PS leaf powder at 2.4 g/d resulted in improving total feed intake and dry matter (DM) digestibility whereas there was reduced protozoal population and methane (CH4) production in cattle.sThe aim of this research was to study the influence of inclusion of Piper sarmentosum (PS) leaf powder on feed efficiency, rumen ecology, protozoal diversity and CH4 emission in cattle. Four male beef cattle (1–1.5 years old) with similar initial body weights (BW) of 150 ± 20.00 kg were randomly assigned to a 4 × 4 Latin squared design. Experimental diets consisted of four levels of PS leaf powder supplementation at 0 (control), 0.6, 1.2 and 2.4 g/head/d, respectively. Dry matter (DM) intake of rice straw and total intake in terms of g/kg BW0.75 differed among treatments and were linearly greatest when PS was added at 2.4 g/head/d (p < 0.05). PS leaf powder did not affect the intake of nutrients and digestibility of organic matter, crude protein, and fibers. Nevertheless, digestibility of DM was found to increase by 4.8% in cattle when added at 2.4 g/head/d as compared to the other control treatments. PS leaf powder did not affect ruminal pH, ruminal temperature, ammonia–nitrogen concentration and blood urea-nitrogen (p > 0.05). The bacterial population was similar across PS levels (p > 0.05). However, the protozoal count was lower in animals fed with the supplement at 2.4 g/head/d (p < 0.05). The PS leaf powder did not affect total volatile fatty acid (VFA) and acetic acid (C2), butyric acid (C4), C2 to propionic acid (C3) and C2 to C3 to C4. However, concentration of C3 at 4 h post feeding and mean value linearly increased with supplementation of PS leaf powder at 2.4 g/d. The 4 h post-feeding and mean values of CH4 concentration were linearly reduced with PS supplementation (p < 0.05). An increase of PS leaf powder at 2.4 g decreased CH4 after feeding 4 h by 21.33% when compared to no PS leaf powder. Thus, supplementation of PS leaf powder at 2.4 g in ruminant feeding is recommended for manipulating rumen efficiency.

Influence of vitamin E on organic matter fermentation, ruminal protein and fatty acid metabolism, protozoa concentrations and transfer of fatty acids.

Vitamin E (Vit. E) is discussed to influence ruminal biohydrogenation. The objective of this study was to investigate the influence of a Vit. E supplementation on rumen fermentation characteristics, ruminal microbial protein synthesis as well as ruminal organic matter fermentation. Furthermore, we aimed to investigate the influence of Vit. E supplementation on short-chain fatty acids (SCFA) and protozoa concentrations in the rumen and, in addition, on transfer rates of middle-chain and long-chain fatty acids into the duodenum in lactating dairy cows. Eight rumen and duodenum fistulated German Holstein cows were assigned to either a group receiving 2,327 IU/d Vit. E (138.6 IU/kg DM DL-α-tocopherylacetate; n=4) or a control group (23.1 IU/kg DM; n=4). Neither ruminal protein synthesis nor organic matter fermentation was influenced by treatment. Vit. E did not act on the concentrations of short-chain fatty acids and protozoa in rumen fluid. Duodenal flow of C13:0 (1.3 versus 0.2 g/d, p=0.014) and iso-C14:0 (1.0 versus 0.5g/d, p=0.050) was higher in the Vit. E group. We observed a trend for higher duodenal flows for C12:0 (1.6 versus 0.9g/d, p=0.095) and anteiso-C15:0 (12.2 versus 8.9g/d, p=0.084). Transfer rate of C12:0 tended to be higher in the Vit. E group (125.61 versus 73.96, p=0.082). No other transfer rates were affected by treatment. Further studies are necessary to investigate the influence of Vit. E on rumen microbiota and their fatty acid production as well as on the impact of different doses of Vit. E supplementation on variables of protein synthesis efficiency.

Reduction of Cryptosporidium, Giardia, and Fecal Indicators by Bardenpho Wastewater Treatment.

Increased demand for water reuse and reclamation accentuates the importance for optimal wastewater treatment to limit protozoa in effluents. Two wastewater treatment plants utilizing advanced Bardenpho were investigated over a 12-month period to determine the incidence and reduction of Cryptosporidium, Giardia, Cyclospora, and fecal indicators. Results were compared to facilities that previously operated in the same geographical area. Protozoa (oo)cysts were concentrated using an electronegative filter and subsequently detected by fluorescent microscopy and/or PCR methods. Cryptosporidium and Giardia were frequently detected in raw sewage, but Cyclospora was not detected in any wastewater samples. Facilities with Bardenpho treatment exhibited higher removals of (oo)cysts than facilities utilizing activated sludge or trickling filters. This was likely due to Bardenpho systems having increased solid wasting rates; however, this mechanism cannot be confirmed as sludge samples were not analyzed. Use of dissolved-air-flotation instead of sedimentation tanks did not result in more efficient removal of (oo)cysts. Concentrations of protozoa were compared with each other, Escherichia coli, somatic coliphage, and viruses (pepper mild mottle virus, Aichi virus 1, adenovirus, and polyomaviruses JC and BK). Although significant correlations were rare, somatic coliphage showed the highest potential as an indicator for the abundance of protozoa in wastewaters.

Occurrence, genotyping, and health risk of Cryptosporidium and Giardia in recreational lakes in Tianjin, China

Cryptosporidium and Giardia are critical parasites in the etiology of diarrhea worldwide, and often cause waterborne outbreaks. The presence of Cryptosporidium and Giardia in recreational lakes was investigated with molecular characterization, and a comprehensive quantitative microbial risk assessment (QMRA) of protozoan infections was performed, considering multiple exposure pathways, differences in age, sex, and disease severity, and the genotypes of the protozoa. Forty-three (82.7%) and 51 (98.1%) water samples were positive for Cryptosporidium oocysts and Giardia cysts, respectively, with average counts of 3.65 oocysts/10 L and 12.58 cysts/10 L, respectively. Six Cryptosporidium species and three Giardia lamblia assemblages were confirmed with molecular analyses. The protozoan concentration was significantly associated with water turbidity, but not with the total coliform numbers. Swimming in the lakes entailed the highest incidence risk of 5.72 × 10−4 per person per year (pppy) (95% confidence interval (CI): 0.03–43.33 × 10−4) for Cryptosporidium and 4.04 × 10−4 pppy (95% CI: 0.01–32.66 × 10−4) for Giardia, whereas wading entailed the lowest risk (2.20 × 10−4 and 1.70 × 10−4 pppy, respectively). The annual burdens attributable to recreational-water-associated cryptosporidiosis and giardiasis were 3.44 (95% CI: 0.04–23.51) and 1.81 (95% CI: 0.01–12.96) disability-adjusted life years per 1,000,000 individuals per year, respectively. Children were more likely to have an individual disease burden than adults, and males were more likely than females. Sensitivity analysis highlighted the great importance of controlling the proportion of exposed individuals and reducing the frequency of exposure. The methodology and results of this study will allow us to better evaluate and reduce the burden of Cryptosporidium and/or Giardia infections associated with recreational water use in China and other countries.

Influence of food, storage temperature, and time on the extracorporeal viability of ruminal fluid of cattle

Rumen fluid obtained from slaughter houses is sometimes used for transfaunation. In this study, we evaluated the ruminal fluid of cattle fed on pasture or pasture and concentrate and slaughtered recently. The rumen fluid samples were stored in a water bath, at ambient temperature, cooled, or frozen and subjected to physical, chemical, and microbiological evaluation for 24 h. The color, consistency, and odor changed primarily in the cooled samples, followed by the ambient temperature and water bath samples. The pH of fresh ruminal fluid was 7.5 in the pasture group and 6.4 in the pasture and concentrate group. The methylene blue reduction time of the fresh ruminal fluid was 2.35 min in the pasture group and 1.86 min in the pasture and concentrate group; the best values were observed in the water bath group. The chloride content was 15.7 mEq/l in the pasture group and 16.3±3.6 mEq/l in the pasture and concentrate group. A predominance of gram-negative bacteria was observed. The concentration of protozoa was 51.5 in the pasture group and 47.5 × 104/ml in the pasture and concentrate group, with a slight predominance of small protozoa; motility was better in the water bath samples than in the ambient temperature, cooled, and frozen samples. The viability of ruminal fluid collected from freshly slaughtered cattle was influenced by food provided in vivo; however, changes as a function of time and storage temperature were more remarkable. The rumen fluid was viable for up to 9 h when stored in the water bath (38°C) and 2 h at ambient temperature, and it was observed to be nonviable when subjected to cooling or freezing.

Strongyle Infection and Gut Microbiota: Profiling of Resistant and Susceptible Horses Over a Grazing Season.

Gastrointestinal strongyles are a major threat to horses' health and welfare. Given that strongyles inhabit the same niche as the gut microbiota, they may interact with each other. These beneficial or detrimental interactions are unknown in horses and could partly explain contrasted susceptibility to infection between individuals. To address these questions, an experimental pasture trial with 20 worm-free female Welsh ponies (10 susceptible (S) and 10 resistant (R) to parasite infection) was implemented for 5 months. Fecal egg counts (FEC), hematological and biochemical data, body weight and gut microbiological composition were studied in each individual after 0, 24, 43, 92 and 132 grazing days. R and S ponies displayed divergent immunological profiles and slight differences in microbiological composition under worm-free conditions. After exposure to natural infection, the predicted R ponies exhibited lower FEC after 92 and 132 grazing days, and maintained higher levels of circulating monocytes and eosinophils, while lymphocytosis persisted in S ponies. Although the overall gut microbiota diversity and structure remained similar during the parasite infection between the two groups, S ponies exhibited a reduction of bacteria such as Ruminococcus, Clostridium XIVa and members of the Lachnospiraceae family, which may have promoted a disruption of mucosal homeostasis at day 92. In line with this hypothesis, an increase in pathobionts such as Pseudomonas and Campylobacter together with changes in several predicted immunological pathways, including pathogen sensing, lipid metabolism, and activation of signal transduction that are critical for the regulation of immune system and energy homeostasis were observed in S relative to R ponies. Moreover, S ponies displayed an increase in protozoan concentrations at day 92, suggesting that strongyles and protozoa may contribute to each other's success in the equine intestines. It could also be that S individuals favor the increase of these carbohydrate-degrading microorganisms to enhance the supply of nutrients needed to fight strongyle infection. Overall, this study provides a foundation to better understand the mechanisms that underpin the relationship between equines and natural strongyle infection. The profiling of horse immune response and gut microbiota should contribute to the development of novel biomarkers for strongyle infection.

Effects of total replacement of soybean meal and corn on ruminal fermentation, volatile fatty acids, protozoa concentration, and gas production

Abstract. The main purpose of this study is to evaluate the effect of total replacement of soybean meal and corn with triticale and faba bean or field pea on rumen fermentation, protozoa counts, and gas production of lactating ewes. A total of 30 Sicilo-Sarde ewes were randomly allocated into three groups and were fed 1.8 kg drymatter of oat hay plus 500 g of one of three concentrates: the first concentrate (CS) was mainly composed of soybean meal, corn, and barley; the second (TFB) was formed by triticale and faba bean; and the third (TFP) was composed of triticale and field pea. The type of concentrate did not affect ruminal pH or ammonia nitrogen concentration (P &gt; 0.05). The individual concentrations of volatile fatty acids showed a significant interaction between the type of concentrate and sampling time (P &lt; 0.05), except for Butyric and Isobutyric acids. Within a post-feeding time, the pattern of evolution of total volatile fatty, acetic, and propionic acids differed significantly at 2 h post feeding (P &lt; 0.05), while butyric and valeric acid changed at 0 and 4 h post feeding. The type of concentrate affected the total number of ciliate protozoa and the Isotricha species (P &lt; 0.05), whereas Entodinium, Ophryoscolex, and Polyplastron were similar among concentrates (P &gt; 0.05). The cumulative gas production from the in vitro fermentation, the time of incubation, and their interaction was affected by concentrate (P &lt; 0.001). The substitution of soybean meal and corn in the concentrate with faba bean or field peas and triticale might maintain rumen parameters of dairy ewes.

Nutritive evaluation and milk quality of including of tomato or olive by-products silages with sunflower oil in the diet of dairy goats

There are worldwide a number of by-products with a strong potential for use in animal feeding to cover purposes as farm economic profitability enhancement or searching of functional food ingredients. Probably fruit and vegetable industry (i.e. olive milling wastes) are amongst the most representative sectors. Two experiments were performed with lactating Murciano-Granadina goats. In Experiment 1 the objective was to assess the effect of the partial replacement of the forage in the diet with olive by-products or tomato silages supplemented with sunflower oil (SFO, 20g/kg DM) on the nutrient utilization, rumen fermentation parameters and microbial abundances, methane production and milk yield and composition. Eighteen goats were distributed in three experimental groups and fed control diet (CD) against diets containing olive by-products silage (OBSD) or tomato surplus silage (TSD). Both OBSD and TSD resulted in higher fat intake and apparent digestibility, higher digestible and metabolizable energy and total purine derivatives excretion. The rest of measured parameters remained unaffected. Diets OBSD and TSD promoted higher milk content in C18:1 trans-11, C20:2 and CLA. Milk fat of goats consuming TSD had the lowest content in C4:0, C6:0 and C8:0, while OBSD produced more total saturated FA (SFA) and less C18:1 n-9 cis and both total mono- and polyunsaturated FA. The acetate concentration was lower in the rumen of animals fed silages, while the butyrate and protozoa concentrations were higher in OBSD. The CH4 production (L/kg DMI) tended to be lower in goats consuming TSD. Experiment 2 was conducted to test the persistence (over 90 days) and in a larger number of animals (n=24) of the effects observed with TSD diet in experiment 1. The TS diet promoted higher DM intake and body weight and lower acetate:propionate ratio than CD, whereas milk yield and composition were not affected. The FA profile was modified by increases in SFA and reducing monounsaturated FA and CLA. It may be concluded that the strategy of partial replacement of conventional forage by local by-products silage combined with SFO supplementation is valuable nutritional strategy in dairy goats since it may improve the animal energy balance while ruminal fermentation, nutrients utilization and milk yield are not compromised.

Effects of bacterial direct-fed microbials on ruminal characteristics, methane emission, and milk fatty acid composition in cows fed high- or low-starch diets

This study investigated the effects of bacterial direct-fed microbials (DFM) on ruminal fermentation and microbial characteristics, methane (CH4) emission, diet digestibility, and milk fatty acid (FA) composition in dairy cows fed diets formulated to induce different ruminal volatile fatty acid (VFA) profiles. Eight ruminally cannulated dairy cows were divided into 2 groups based on parity, days in milk, milk production, and body weight. Cows in each group were fed either a high-starch (38%, HS) or a low-starch (2%, LS) diet in a 55:45 forage-to-concentrate ratio on a dry matter (DM) basis. For each diet, cows were randomly assigned to 1 of 4 treatments in a Latin square design of (1) control (CON); (2) Propionibacterium P63 (P63); (3) P63 plus Lactobacillus plantarum 115 (P63+Lp); (4) P63 plus Lactobacillus rhamnosus 32 (P63+Lr). Strains of DFM were administered at 1010 cfu/d. Methane emission (using the sulfur hexafluoride tracer technique), total-tract digestibility, dry matter intake, and milk production and composition were quantified in wk 3. Ruminal fermentation and microbial characteristics were measured in wk 4. Data were analyzed using the mixed procedure of SAS (SAS Institute Inc., Cary, NC). The 2 diets induced different ruminal VFA profiles, with a greater proportion of propionate at the expense of acetate and butyrate for the HS diet. Greater concentrations of total bacteria and selected bacterial species of methanogenic Archaea were reported for the HS diet, whereas the protozoa concentration in HS decreased. For both diets, bacterial DFM supplementation raised ruminal pH (+0.18 pH units, on average) compared with CON. Irrespective of diet, P63+Lp and P63+Lr increased ruminal cellulase activity (3.8-fold, on average) compared with CON, but this effect was not associated with variations in ruminal microbial numbers. Irrespective of diet, no effect of bacterial DFM on ruminal VFA was observed. For the LS diet, supplementing cows with P63+Lr tended to decrease CH4 emission (26.5%, on average, when expressed per kilogram of milk or 4% fat-corrected milk). Only P63 supplementation to cows fed the HS diet affected the concentration of some milk FA, such as cis isomers of 18:1 and intermediates of ruminal biohydrogenation of polyunsaturated FA. Overall, bacterial DFM could be useful to stabilize ruminal pH. Their effects on CH4 production mitigation and milk FA profile depended on DFM strain and diet and should be confirmed under a greater variation of dietary conditions.

Tea saponin reduced methanogenesis in vitro but increased methane yield in lactating dairy cows

The effect of tea saponin supplementation in the ruminant diet on methane emissions, rumen fermentation, and digestive processes is still under debate. The objective of this study was to assess the effect of this plant extract on methanogenesis, total-tract digestibility, and lactating performances of dairy cows. The work included 2 independent and successive experiments. First, the effect of 7 tea saponin doses (from 0 to 0.50 g/L) on methane emissions and protozoa concentrations was tested in 2 repeated in vitro batch culture incubations using bovine rumen contents as inoculum and a cereal mixture as substrate. After 18 h of incubation, total gas production and composition as well as rumen fermentation parameters and protozoa concentration were analyzed. Increasing dosage of the plant extract reduced methane production and protozoa concentration, with a maximum reduction of 29% for CH4 (mL/g of substrate) and 51% for protozoa (105/mL). Tea saponin did not affect volatile fatty acids concentration, but marginally decreased total gas production by 5% at the highest dose. Second, a 2-period crossover design experiment was carried out with 8 lactating dairy cows fed a basal diet (54% corn silage, 6% hay, and 40% pelleted concentrates on a dry matter basis) without (control) or with 0.52% tea saponin (TSP). Each experimental period lasted 5 wk. Animals were fed ad libitum during the first 3 wk of the period (wk 1, 2, and 3) and restricted (95% of ad libitum intake) during the last 2 wk (wk 4 and 5). Intake and milk production were recorded daily. Methane emissions were quantified using open chambers (2 d, wk 4). Total-tract digestibility and nitrogen balance were determined from total feces and urine collected separately (5 d, wk 5). Rumen fermentation parameters and protozoa concentration were analyzed from samples taken after morning feeding (1 d, wk 5). Milk production, dry matter intake, and feed efficiency were reduced with TSP (-18, -12, and -8%, respectively). As daily methane production (g/d) was not affected, methane emissions (g/kg of dry matter intake) increased by 14% with TSP. Total-tract digestibility and nitrogen balance were similar between diets, except for acid detergent fiber digestibility, which tended to be improved with TSP (+4 percentage units). Rumen fermentation parameters and protozoa concentration were relatively unchanged by diets. Under the conditions of this experiment, tea saponin is not efficient to reduce methane emissions from dairy cows.