Connection of spectral pattern of carbohydrate molecular structure to alteration of nutritional properties of by-products from coffee processing affected by microorganism fermentation.

Objective: The objective of this study was to determine internal structure spectral profile of by-products from coffee processing that were affected by added-microorganism fermentation duration in relation to truly absorbed feed nutrient supply in ruminant system. Methods: The by-products from coffee processing were fermented using commercial fermentation product, consisting of various microorganisms: for 0 (control), 7, 14, 21, and 28 days. In this study, the association and correlation of carbohydrate-related spectral profiles with chemical and nutritional properties (chemical composition, total digestible nutrient, bioenergy values, carbohydrate sub-fractions and predicted degradation and digestion parameters as well as milk value of feed. The vibrational spectra of coffee by-products samples after fermentation for 0 (control), 7, 14, 21, and 28 days were determined using a JASCO FT/IR-4200 spectroscopy coupled with accessory of Attenuated total reflectance (ATR). The molecular spectral analyses with univariate approach were conducted with the OMNIC 7.3 software. Results: Molecular spectral analysis parameters in fermented and non-fermented by-products from coffee processing included Structural carbohydrate, Cellulosic compounds, Non-structural carbohydrates, Lignin compound, CH-bending, structural carbohydrate peak1, Structural carbohydrate peak2, Structural carbohydrate peak3, Hemicellulosic compound, Non-structural carbohydrate peak1, non-structural carbohydrate peak2, non-structural carbohydrate peak3. The study results show that added-microorganism fermentation induced chemical and nutritional changes of coffee by-products including carbohydrate chemical composition profiles, bioenergy value, feed milk value, carbohydrate subfractions, estimated degradable and undegradable fractions in the rumen, and intestinal digested nutrient supply in ruminant system. Conclusion: In conclusion, carbohydrate nutrition value changes by added-microorganism fermentation duration were in an agreement with the change of their spectral profile in the coffee by-products. The studies show that the vibrational ATR-FT/IR spectroscopic technique could be applied as a rapid analytical tool to evaluate fermented by-products and connect with truly digestible carbohydrate supply in ruminant system.

Foot-and-mouth disease seroprevalence is higher in young female crossbred sheep and goats compared to their counterparts in the border area between Pakistan and Afghanistan.

OBJECTIVE: Foot-and-mouth disease (FMD) is a highly contagious disease in ruminants that causes significant economic losses worldwide. However, the prevalence of FMD virus (FMDV) in small ruminants has been overlooked in Pakistan. This study aimed to determine the prevalence of FMD in sheep and goats in the border area between Pakistan and Afghanistan. ANIMALS: 800 sheep and goats belongs to age groups of 6 month to > 2 years. METHODS: A total of 800 serum samples were collected from sheep (n = 424) and goats (n = 376) and subjected to structural protein (SP) and 3ABC non-SP (NSP) ELISAs for the detection of antibodies against SP and NSP of the FMDV. RESULTS: For NSP, 340/800 (42.5%) of samples were positive, while SP analysis revealed that serotype O (44.5%) was the most common in sheep and goats, followed by Asia-1 (42%) and A (32%) serotypes. Sheep (39%; 95% CI, 34 to 44) had a higher (P < .05) prevalence of FMD than goats (46%; 95% CI, 41 to 51). Statistically significant (P < .05) differences in the seroprevalence of FMD-SP and FMD-NSPs were observed between various agencies (areas) of the study area. Risk factors such as age, sex, breed, season, flock size, body condition, animal movement, and production system were significantly (P < .05) associated with FMDV prevalence. CONCLUSIONS: This study showed that FMD is highly prevalent in sheep and goats in the border areas of Pakistan and Afghanistan. Therefore, outbreak investigation teams should be arranged at the border level to develop FMD risk-based surveillance and control plans for small ruminants in order to mitigate infection risks.

Understanding the Diversity and Roles of the Ruminal Microbiome.

The importance of ruminal microbiota in ruminants is emphasized, not only as a special symbiotic relationship with ruminants but also as an interactive and dynamic ecosystem established by the metabolites of various rumen microorganisms. Rumen microbial community is essential for life maintenance and production as they help decompose and utilize fiber that is difficult to digest, supplying about 70% of the energy needed by the host and 60-85% of the amino acids that reach the small intestine. Bacteria are the most abundant in the rumen, but protozoa, which are relatively large, account for 40-50% of the total microorganisms. However, the composition of these ruminal microbiota is not conserved or constant throughout life and is greatly influenced by the host. It is known that the initial colonization of calves immediately after birth is mainly influenced by the mother, and later changes depending on various factors such as diet, age, gender and breed. The initial rumen microbial community contains aerobic and facultative anaerobic bacteria due to the presence of oxygen, but as age increases, a hypoxic environment is created inside the rumen, and anaerobic bacteria become dominant in the rumen microbial community. As calves grow, taxonomic diversity increases, especially as they begin to consume solid food. Understanding the factors affecting the rumen microbial community and their effects and changes can lead to the early development and stabilization of the microbial community through the control of rumen microorganisms, and is expected to ultimately help improve host productivity and efficiency.

Epidemiology of Digital Dermatitis in Western Canadian Feedlot Cattle.

Digital dermatitis (DD) is an emerging disease in feedlot cattle. Our objective was to identify animal- and feedlot-level risk factors for DD by analyzing individual animal health records (n = 1,209,883) and feedlot-level records from western Canadian feedlots (n = 28) between 2014 and 2018, inclusive. The risk of a DD diagnosis was higher (incidence rate ratio (IRR) = 2.08, 95% CI 1.52 to 2.86) in cattle sourced from confined background operations (CB) versus cattle sourced from auction markets (AM). Conversely, ranch direct (RD) cattle were (IRR = 0.02, 95% CI 0.04 to 0.30) lower risk than AM cattle of being diagnosed with DD. The risk of being diagnosed with DD was higher in females than in males. The magnitude of the risk in females over males was influenced by annual DD incidence in low morbidity years (2014, 2017, and 2018) (IRR = 2.02, 95% CI 1.27 to 3.19), medium morbidity years (2016) (IRR = 2.95, 95% CI 1.64 to 5.33), and high morbidity years (2015) (IRR = 5.41, 95% CI 3.27 to 8.95). At the feedlot-level, the risk of a diagnosis of DD was lower in small capacity (SCF) versus large capacity feedlots (LCF) (IRR = 0.24, 95% CI 0.05 to 0.76). Future research should focus on identifying factors that may propagate disease transmission between cattle of different sexes and from different acquisition sources.

Use of irradiated chitosan as a matrix for slow-release urea and in vitro fermentation characteristics of slow-release urea supplementation in ruminant rations.

Background and Aim: Irradiated chitosan can be used as a matrix for slow-release urea (SRU) production. This study aimed to (1) determine the optimal formulation of irradiated chitosan matrix for controlling nitrogen release and (2) evaluate the characteristics of SRU in vitro fermentation based on irradiated chitosan as a feed supplement. Materials and Methods: In the first phase of the investigation, four chitosan-based SRU formulations with varying amounts of acrylamide (3 and 5 g) and gamma irradiation (5 and 10 kGy) were evaluated. Scanning electron microscopy, Fourier transform mid-infrared spectroscopy, and ammonia release characteristics were used to observe morphological, functional group, and ammonia release characteristics. In the second phase of research, the most effective SRU formulation was utilized as a supplement to ruminant rations based on rice straw, sorghum straw, and alfalfa. Gas production, rumen fermentation characteristics, and methane gas production were observed in vitro. Results: On the basis of surface image analysis, the four SRU formulas generate a similar appearance. Compared with untreated urea, the SRU3 formula reduced the percentage of ammonia emission by 12.85%-27.64% after 24 h of incubation (p = 0.05), as determined by the first phase study. SRU3 became the basis for the second testing phase. The addition of SRU3 did not affect the optimal gas production in vitro. SRU3 treatment produced less gas than Optigen® treatment (p = 0.05). With regard to rumen fermentation and digestibility, Optigen® yielded better results than SRU3 (p = 0.05). However, the treatment with SRU3 resulted in reduced methane production compared to that in the control (p = 0.05). Conclusion: Irradiated chitosan as an SRU matrix may control the release of ammonia in the rumen medium. The SRU3 formulation is the most effective. The addition of SRU to rice straw-based rations reduces methane production without affecting in vitro digestibility.

A Combined Approach for the Characterization of Small Ruminant Lentivirus Strains Circulating in the Islands and Mainland of Greece.

Small ruminant lentiviruses are a group of viruses infecting goat and sheep worldwide. These viruses exhibit an extraordinary degree of genetic and antigenic variability that severely influence in vivo and in vitro features, as well as diagnostic test results. Small ruminant farming is the most important animal farming business in Greece, with a high impact on the Greek primary economy. Although SRLV infection and its impact on animal production are well established in the country, little is known about the circulating SRLV strains and their prevalence. The aim of this study was to characterize SRLVs circulating in Greece with a combined serological and molecular approach, using the bulk milk matrix collected from 60 farms in different municipalities. This study allowed us to estimate a seroprevalence of around 52% at the herd level. The B1, B2 and A3 subtypes and a novel A viral cluster were identified. Moreover, the amplicon sequencing method allowed us to identify more than one viral subtype in a sample. These results again confirm the high variability of these viruses and highlight the importance of the constant monitoring of viral evolution, in particular in antigens of diagnostic interest.

Corrugated intimal surface of the ovine aorta: when physiology resembles pathology.

The aortic lumen in healthy animals is characterized by a smooth, whitish surface, but sheep have macroscopic corrugation of the intimal surface in the thoracic aorta (TA). Our aim was to determine if this finding was pathological or physiological. Thirteen sheep aortas were included in this work together with aortas from cattle (n = 3), a goat (n = 1), horses (n = 4), dogs (n = 2), rabbits (n = 2) and a pig (n = 1). A corrugated intimal surface in the TA was seen in all the sheep and the goat but was less evident in the cattle. Histologically, in sheep the TA intimal surface was seen to have multifocal bulging areas that protruded into the lumen. The outer half of the tunica media had numerous, randomly distributed muscle islands that disrupted the arrangement of the elastic lamella, displacing them towards the lumen. We conclude that the intimal corrugation of the TA in sheep is physiological and must not be misinterpreted as pathological.

Prevalence and risk factors of fasciolosis in a bovine population from farms in Taiping, Malaysia.

Fasciolosis is a zoonotic disease, considered an emerging neglected tropical disease threatening ruminant productivity and causing economic losses. Controlling fasciolosis is challenging due to the complex life cycle of Fasciola, which involves snail intermediate hosts. The high rainfall status in Taiping makes it an optimal region for snail abundance, which increases the opportunity to complete Fasciola's life cycle. Previous studies showed that liver condemnation caused by fasciolosis was highly prevalent in the Taiping abattoir compared to other investigated main abattoirs of Peninsular Malaysia. Therefore, the present study determined the prevalence of bovine fasciolosis and risk factors in farms from Larut and Matang (Taiping), Malaysia. Sampling was carried out from February until August 2020. In this cross-sectional study, a total of 371 fecal samples from bovines (dairy cattle, beef cattle, buffalo) were examined from 23 farms selected based on location, farmer consent, and history of anthelmintic usage. Animal's intrinsic and farm management details were recorded, and interview sessions were conducted with farmers to collect information on the potential risk factors. Individual fecal samples were examined for the presence of Fasciola egg using Flukefinder® sedimentation. There was moderate prevalence of bovine fasciolosis in Taiping (36.9%, n = 137/371). Significant risk factors (p < 0.05) were observed, which include buffalo group (OR = 9.5, 95% CI: 9.44-9.55), age of >3 years (OR = 5.5, 95% CI: 5.43-5.57), thinner animals with body condition score of 1 to 4 (OR = 1.2-14.9, 95% CI: 1.09-15.08), and larger grazing area (OR = 1.3, 95% CI: 1.30-1.31). Additional risk factors include the presence of more than one ruminant species in the same farm (OR = 2.0-2.1, 95% CI: 2.00-2.22), extensive housing system (OR = 4.0, 95% CI: 3.77-4.23), farm age (OR = 1.2, 95% CI: 1.20-1.21), and also co-infection with Paramphistomes (OR = 1.4, 95% CI: 1.10-1.71). The present study underscores the importance of local bovine fasciolosis epidemiology, which could be used to conduct future veterinary and public health programmes to inform effective parasitic management aimed at reducing the prevalence of fasciolosis.

Role of Silver Nanoparticles for the Control of Anthelmintic Resistance in Small and Large Ruminants.

Helminths are considered a significant threat to the livestock industry, as they cause substantial economic losses in small and large ruminant farming. Their morbidity and mortality rates are also increasing day by day as they have zoonotic importance. Anthelmintic drugs have been used for controlling these parasites; unfortunately, due to the development of resistance of these drugs in helminths (parasites), especially in three major classes like benzimidazoles, nicotinic agonists, and macrocyclic lactones, their use is becoming very low. Although new anthelmintics are being developed, the process is time-consuming and costly. As a result, nanoparticles are being explored as an alternative to anthelmintics. Nanoparticles enhance drug effectiveness, drug delivery, and target specificity and have no resistance against parasites. Different types of nanoparticles are used, such as organic (chitosan) and inorganic (gold, silver, zinc oxide, iron oxide, and nickel oxide). One of them, silver nanoparticles (AgNPs), has unique properties in various fields, especially parasitology. AgNPs are synthesized from three primary methods: physical, chemical, and biological. Their primary mechanism of action is causing stress through the production of ROS that destroys cells, organs, proteins, and DNA parasites. The present review is about AgNPs, their mode of action, and their role in controlling anthelmintic resistance against small and large ruminants.

Quantifying the Impact of Different Dietary Rumen Modulating Strategies on Enteric Methane Emission and Productivity in Ruminant Livestock: A Meta-Analysis.

A meta-analysis was conducted with an aim to quantify the beneficial effects of nine different dietary rumen modulating strategies which includes: the use of plant-based bioactive compounds (saponin, tannins, oils, and ether extract), feed additives (nitrate, biochar, seaweed, and 3-nitroxy propanol), and diet manipulation (concentrate feeding) on rumen fermentation, enteric methane (CH4) production (g/day), CH4 yield (g/kg dry matter intake) and CH4 emission intensity (g/kg meat or milk), and production performance parameters (the average daily gain, milk yield and milk quality) of ruminant livestock. The dataset was constructed by compiling global data from 110 refereed publications on in vivo studies conducted in ruminants from 2005 to 2023 and anlayzed using a meta-analytical approach.. Of these dietary rumen manipulation strategies, saponin and biochar reduced CH4 production on average by 21%. Equally, CH4 yield was reduced by 15% on average in response to nitrate, oils, and 3-nitroxy propanol (3-NOP). In dairy ruminants, nitrate, oils, and 3-NOP reduced the intensity of CH4 emission (CH4 in g/kg milk) on average by 28.7%. Tannins and 3-NOP increased on average ruminal propionate and butyrate while reducing the acetate:propionate (A:P) ratio by 12%, 13.5% and 13%, respectively. Oils increased propionate by 2% while reducing butyrate and the A:P ratio by 2.9% and 3.8%, respectively. Use of 3-NOP increased the production of milk fat (g/kg DMI) by 15% whereas oils improved the yield of milk fat and protein (kg/d) by 16% and 20%, respectively. On the other hand, concentrate feeding improved dry matter intake and milk yield (g/kg DMI) by 23.4% and 19%, respectively. However, feed efficiency was not affected by any of the dietary rumen modulating strategies. Generally, the use of nitrate, saponin, oils, biochar and 3-NOP were effective as CH4 mitigating strategies, and specifically oils and 3-NOP provided a co-benefit of improving production parameters in ruminant livestock. Equally concentrate feeding improved production parameters in ruminant livestock without any significant effect on enteric methane emission. Therefore, it is advisable to refine further these strategies through life cycle assessment or modelling approaches to accurately capture their influence on farm-scale production, profitability and net greenhouse gas emissions. The adoption of the most viable, region-specific strategies should be based on factors such as the availability and cost of the strategy in the region, the specific goals to be achieved, and the cost-benefit ratio associated with implementing these strategies in ruminant livestock production systems.

Dietary carbohydrate sources differently prime the microbial ecosystem but not the epithelial gene expression profile along the complete gut of young calves.

BACKGROUND: Recent data indicated similar growth performance of young calves fed solely high-quality hay instead of a starter diet based on starchy ingredients. Yet, providing exclusively such distinct carbohydrate sources during early life might specifically prime the microbiota and gene expression along the gut of young calves, which remains to be explored. We investigated the effects of starter diets differing in carbohydrate composition, that is medium- or high-quality hay and without or with 70% concentrate supplementation (on fresh matter basis), across the gastrointestinal tract (GIT) of weaned Holstein calves (100 ± 4 days of age) using 16 S rRNA gene sequencing and analyses of short-chain fatty acids and host epithelial gene expressions. RESULTS: The concentrate supplementation drastically decreased microbial diversity throughout the gut, which was also true to a much lesser extent for high-quality hay when compared to medium-quality hay in the foregut. Similarly, the factor concentrate strongly shaped the diet-associated common core microbiota, which was substantially more uniform along the gut with concentrate supplementation. The fermentation profile shifted towards less acetate but more propionate with concentrate supplementation in almost all gut sections, corresponding with higher abundances of starch-utilizing bacteria, while major fibrolytic clusters declined. Noteworthy, the n-butyrate proportion decreased in the rumen and increased in the colon with concentrate, showing an opposite, gut site-dependent effect. Both dietary factors modestly influenced the host epithelial gene expression. CONCLUSIONS: Concentrate supplementation clearly primed the microbial ecosystem on a starch-targeted fermentation with characteristic genera occupying this niche along the entire GIT of calves, whereas the microbial differentiation due to hay quality was less distinct. Overall, changes in the microbial ecosystem were only marginally reflected in the targeted transcriptional profile of the host epithelium.

Physical Pretreatments of Cassava Chips Influenced Chemical Composition, Physicochemical Properties, and In Vitro Digestibility in Animal Models.

Physical pretreatment procedures can significantly influence the quality of food and feed raw materials. To increase the ability to be digested in animals, cassava chips were pretreated by four alternative methods (extrusion, microwave irradiation, gamma irradiation, or NaOH hydrolysis), and then the chemical composition, physicochemical properties, and in vitro digestibility of the pretreated samples were assessed and compared with unprocessed cassava chips (control). The chemical compositions (crude protein, ether extract, neutral detergent fiber, acid detergent fiber, ash, non-fiber carbohydrate, and gross energy) were significantly altered due to the pretreatment methods (p < 0.05). The nutritive profile was qualitatively changed when assessed through Fourier-transform infrared spectroscopy. Some physicochemical properties in association with enzymatic hydrolysis, which include pH, water solubility, water absorption capacity, thermal properties (differential scanning calorimetry), diffraction pattern (X-ray diffractometry), and microstructure (scanning electron microscopy), were significantly changed. In vitro carbohydrate digestibility based on digestive enzyme extracts from Nile tilapia (Oreochromis niloticus) suggests the microwaving method for cassava chips preparation, while microwaving, followed by extrusion or gamma irradiation, was suggested for broiler (Gallus gallus domesticus). There were no differences in the pepsin-cellulase digestibility values tested for the ruminant model. These findings suggest the use of pretreated cassava chips in animal feeding.

Behavioural Factors Influencing the Intention to Adopt Sheep Scab Control Measures in Northern Ireland.

Sheep scab, caused by infestation with the ectoparasitic mite Psoroptes ovis, is an endemic disease in the Northern Ireland (NI) sheep flock and constitutes significant economic and welfare burdens for the NI farming industry. Despite its endemic nature, historically, little research has been undertaken to support the control of the disease in NI. This study offers the first attempt to analyse the psychological and behavioural factors influencing farmers' intentions to implement effective sheep scab control measures in NI. To achieve our objective, quantitative data from a sample of 126 sheep farmers were statistically analysed using an extended theory of planned behaviour approach in an ordered logistic regression modelling framework. Our analyses showed that sheep scab remains an issue of concern in Northern Ireland. The attitudes of the farmers, as well as perceived behavioural control, emotional effect, membership of Business Development Groups (BDGs), and higher education qualifications, were found to be statistically significant factors influencing farmers' intentions to adopt sheep scab control measures. This study provides a solid foundation for how to promote behavioural changes among sheep farmers to improve their ability to implement effective disease control measures, helping to tackle this challenging disease in a more sustainable way in the future.

Vimentin inhibits peste des petits ruminants virus replication by interaction with nucleocapsid protein.

The Peste des petits ruminant virus (PPRV) is a member of the Paramyxoviridae family and is classified into the genus Measles virus. PPRV predominantly infects small ruminants, leading to mortality rates of nearly 100%, which have caused significant economic losses in developing countries. Host proteins are important in virus replication, but the PPRV nucleocapsid (N) protein-host interacting partners for regulating PPRV replication remain unclear. The present study confirmed the interaction between PPRV-N and the host protein vimentin by co-immunoprecipitation and co-localization experiments. Overexpression of vimentin suppressed PPRV replication, whereas vimentin knockdown had the opposite effect. Mechanistically, N was subjected to degradation via the ubiquitin/proteasome pathway, where vimentin recruits the E3 ubiquitin ligase NEDD4L to fulfill N-ubiquitination, resulting in the degradation of the N protein. These findings suggest that the host protein vimentin and E3 ubiquitin ligase NEDD4L have an anti-PPRV effect.

Molecular characterization and haplotypes of hydatid cyst isolates collected from humans and ruminants in Setif Province (northeast of Algeria) based on mitochondrial cytochrome C oxidase subunit 1 (mt-CO1) gene sequences.

Cystic echinococcosis (CE) is a disease that can be transmitted from animals to humans, caused by the metacestode of Echinococcus granulosus. The disease has significant health and economic impacts worldwide, particularly in endemic areas. The study aimed to evaluate the prevalence of hydatid cysts in ruminants (cattle and sheep) (n = 2060) from the Setif Province of Algeria using microscopy. The results showed that hydatid cysts were detected in 9.6% (198/2060) of ruminants, with a higher prevalence in cattle (16.8%; 56/333) compared to sheep (8.2%; 142/1727). Molecular techniques were used to analyze a subset of animals consisting of 30 sheep and 4 cattle. Specifically, a fragment of the mitochondrial cytochrome c oxidase subunit 1 (mt-CO1) gene was sequenced and compared to sequences from seven humans from the same region. The results indicated that all isolates were identified as E. granulosus sensu stricto. Haplotype analysis identified 19 E. granulosus s.s. haplotypes arranged like a star, with the dominant haplotype (Hap04) at the center. Hap04 has been assigned a total of 17 positives, including positives from sheep, cattle, and two humans. This study is noteworthy for being the first to use a molecular approach to human and ruminant echinococcosis in Setif, a significant breeding region in Algeria.

Complete genome sequence of Methanosphaera sp. ISO3-F5, a rumen methylotrophic methanogen.

Methanosphaera spp. are methylotrophic methanogenic archaea and members of the order Methanobacteriales with few cultured representatives. Methanosphaera sp. ISO3-F5 was isolated from sheep rumen contents in New Zealand. Here, we report its complete genome, consisting of a large chromosome and a megaplasmid (GenBank accession numbers CP118753 and CP118754, respectively).

Phylogenetic analysis of small ruminant lentiviruses in Mongolian sheep supports an ancient east-west split for the genotype A.

The ovine maedi-visna virus (MVV) and caprine arthritis-encephalitis virus (CAEV) are small ruminant lentiviruses (SRLVs) with striking genetic and structural similarities. The presence of SRLV in Mongolian sheep and goats was serologically demonstrated more than a decade ago; however, the viral genotype remains unknown. In total, 329 blood samples were collected from two sheep breeds (i.e., Khalkha and Sumber) in Tov, Govisumber, Arkhangay, Dornogovi, Zavkhan, and Sukhbaatar provinces, Mongolia. Serological and phylogenetic analyses were performed regardless of any apparent clinical signs, although most of the animals appeared healthy. All sheep in three of the six provinces were seronegative, whereas the seroprevalence in the Tov, Govisumber, and Zavkhan provinces averaged 7.9%. Genomic DNA from seropositive animals was tested using hemi-nested polymerase chain reaction, and sub-genomic SRLV sequences were determined from nine samples. Mongolian SRLV sequences clustered within the divergent subtype A22, which was previously found only in Fertile Crescent regions, including Lebanon, Jordan, and Iran, where the first sheep-domestication (Ovis aries) occurred. According to the phylogenetic analysis, genotype A has two ancestors from the ancient Fertile Crescent: (1) Turkish strains and (2) Iranian, Jordanian, and Lebanese strains. The first ancestor spread westward, whereas the second spread eastward, ultimately reaching Mongolia.

Sheep fatigue during transport: Lost in translation?

Although sheep are commonly transported long distances, and sheep welfare during transport is a topic of research and policy discussion, the subject of their fatigue during transport has been under-researched. The current qualitative study, focused on the EU and UK, aimed to critically analyse stakeholder views on issues relating to sheep fatigue, including behavioural indications of fatigue, the interplay between fatigue and other factors, and the practicalities of identifying fatigue in commercial transport conditions. Insight into stakeholder perceptions of these issues could contribute to the body of knowledge regarding sheep fatigue during transport, potentially playing a part in future efforts to improve fatigue understanding and detection. Eighteen experts from different stakeholder groups were interviewed. Reflexive thematic analysis of interview data yielded four themes and three sub-themes. The first theme, "Let's anthropomorphise it a little bit", underscores the pervasiveness of anthropomorphism and suggests using it in a conscious and deliberate way to drive stakeholder engagement and policy change. The second theme, "We think that they're like we are and they're not", cautions against wholesale transfer of human experiences to animals. The third theme, 'See the whole animal', advocates using Qualitative Behaviour Analysis (QBA), proven reliable in other contexts, to deepen and enrich our current understanding of fatigue. The fourth theme, 'Fatigue "never comes up"', highlights the fact that fatigue is rarely if ever discussed in the context of sheep transport. These themes suggest several avenues for future research, including developing QBA-based assessments for fatigue to improve welfare during transport.

Optimization of volatile organic compounds sampling from dairy cow exhaled breath using polymer-based solid-phase extraction cartridges for gas chromatographic analysis.

We explored appropriate technical setups for the detection of volatile organic compounds (VOCs) from exhaled cow breath by comparing six different polymer-based solid-phase extraction (SPE) cartridges currently on the market for gas chromatography/mass spectrometry (GC-MS) screening. Exhaled breath was sampled at a single timepoint from five lactating dairy cows using six different SPE cartridges (Bond Elut ENV (ENV); Chromabond HRX (HRX); Chromabond HRP (HRP); Chromabond HLB (HLB); Chromabond HR-XCW (XCW) and Chromabond HR-XAW (XAW)). The trapped VOCs were analyzed by dynamic headspace vacuum in-tube extraction GC-MS (DHS-V-ITEX-GC-MS). Depending on the SPE cartridge, we detected 1174-1312 VOCs per cartridge. Most VOCs were alkenes, alkanes, esters, ketones, alcohols, aldehydes, amines, nitriles, ethers, amides, carboxylic acids, alkynes, azoles, terpenes, pyridines, or sulfur-containing compounds. The six SPE cartridges differed in their specificity for the chemical compounds, with the XAW cartridge showing the best specificity for ketones. The greatest differences between the tested SPE cartridges appeared in the detection of specific VOCs. In total, 176 different VOCs were detected with a match factor >80%. The greatest number of specific VOCs was captured by XAW (149), followed by ENV (118), HLB (117), HRP (115), HRX (114), and XCW (114). We conclude that the tested SPE cartridges are suitable for VOC sampling from exhaled cow breath, but the SPE cartridge choice enormously affects the detected chemical groups and the number of detected VOCs. Therefore, an appropriate SPE adsorbent cartridge should be selected according to our proposed inclusion criteria. For targeted metabolomics approaches, the SPE cartridge choice depends on the VOCs or chemical compound groups of interest based on our provided VOC list. For untargeted approaches without information on the animals' metabolic condition, we suggest using multi-sorbent SPE cartridges or multiple cartridges per animal.

Rumen degradable protein levels on fatty acid profile of rumen bacteria and milk fat of dairy cows supplemented with calcium salts of fatty acids.

This study aimed to evaluate the effects of calcium salts of fatty acids (CSFA) from soybean oil in diets with different levels of rumen degradable protein (RDP) on bio-hydrogenation extent, and fatty acid (FA) profile intake, omasal digesta, rumen bacteria, and milk fat. Eight Holstein lactating cows were used in a replicated 4 × 4 Latin square design. Treatments were arranged in a 2 × 2 factorial design with two CSFA levels (0 or 33.2 g/kg DM of CSFA) and two RDP levels (98.0 or 110 g/kg DM). There was RDP and CSFA interaction effect on the omasal flow of total FA and some specific FA. Only in -CSFA diets, the higher RDP level increased omasal flow of total FA. Dietary RDP levels did not affect the FA profile of bacteria and milk fat. Feeding CSFA reduced or tended to reduce the bacterial proportion of C15:0, C16:0, C16:1, C17:0, and C18:0 FA and decreased the concentrations of short- and medium-chain FA (<18C) and increased the concentrations of unsaturated and long-chain FA (≥18C) in milk fat. Feeding CSFA of soybean oil increases the intake and omasal flow of FA and augments unsaturated FA content in bacteria pellets and milk fat.