Domestic Livestock Species Research Articles

A point of view on genetic resources and plant breeding

Biodiversity is the set of living organisms that make up our battered planet and their interactions with the environment. It is made up of the Kingdoms: Animalia, Plantae, Fungi, Protista, and Monera. It provides the energy resources necessary to sustain the planet and our civilization; it is the basis of agriculture, source of all crops, and species of domestic livestock since the beginning of human civilization.[...]

Maternal nutrition and developmental programming of offspring.

Developmental programming is the concept that 'stressors' during development (i.e. pregnancy, the perinatal period and infancy) can cause long-term changes in gene expression, leading to altered organ structure and function. Such long-term changes are associated with an increased risk of a host of chronic pathologies, or non-communicable diseases including abnormal growth and body composition, behavioural or cognitive dysfunction, metabolic abnormalities, and cardiovascular, gastro-intestinal, immune, musculoskeletal and reproductive dysfunction. Maternal nutrition during the periconceptual period, pregnancy and postnatally can have profound influences on the developmental program. Animal models, including domestic livestock species, have been important for defining the mechanisms and consequences of developmental programming. One of the important observations is that maternal nutritional status and other maternal stressors (e.g. environmental temperature, high altitude, maternal age and breed, multiple fetuses, etc.) early in pregnancy and even periconceptually can affect not only embryonic/fetal development but also placental development. Indeed, altered placental function may underlie the effects of many maternal stressors on fetal growth and development. We suggest that future directions should focus on the consequences of developmental programming during the offspring's life course and for subsequent generations. Other important future directions include evaluating interventions, such as strategic dietary supplementation, and also determining how we can take advantage of the positive, adaptive aspects of developmental programming.

363 Relationship Between Parasite Load and Performance in Goat Kids at Weaning and Early Post Weaning

Abstract Goats are small ruminants that have enormous potential to boost the economy and be a major source of income especially to marginal farmers and landless laborers. A very common and economically important condition, which affects domestic livestock species worldwide, is Gastrointestinal (GI) parasitism. Haemonchus contortus is the gastrointestinal parasite most common in small ruminants. To better understand the impact of GI parasitism, a study was conducted to research the relationship between parasite load and performance in goat kids at weaning and 60 d post-weaning. Fecal samples were collected at weaning and 60 d post-weaning and egg counts were performed using the modified McMasters technique. Data collected included weights, Fecal Egg Count at weaning, and at 60 d post-weaning. The average fecal egg count at weaning was 830 eggs per gram and 1103 eggs per gram 60 d post-weaning for H. contortus. Egg count at weaning was significantly related to weight at both weaning and 60 d post-weaning (P < .0001 and P = .0003 respectively), but not to ADG (P > .05). Fecal egg count at 60 d post-weaning was not significantly related to either weight or ADG (P > .05). Egg counts at weaning and 60 days post-weaning were not related (P = .0708). The results indicate that early parasite infection can influence animal weights. The lack of a significant correlation between egg counts at weaning and 60 d post weaning indicates a need to evaluate parasite load at later ages in kids when selecting for resistance.

Pigs in the Neolithic of the Eastern Fertile Crescent: New evidence from Pre‐Pottery Neolithic Bestansur and Shimshara, Iraqi Kurdistan (7800–7100 BC)

AbstractPigs are one of the earliest domesticated livestock species, first domesticated at least 10,000 years ago. The domestication of wild boar, including associated morphological changes, is a long process over several millennia. Across Southwest Asia, management, domestication, and the adaption of the different livestock species was a highly localized process, influenced by both cultural and environmental factors.This paper explores the size, age, and diet of the suids of the Pre‐Pottery Neolithic sites of Bestansur and Shimshara in order to further our understanding of the origins of suid management in the Neolithic Eastern Fertile Crescent. Our data suggest that the relationship between wild boar and humans was more complex than a strict hunter–prey relationship. This study demonstrates that the Neolithic in the Zagros was not uniform in the adaption and exploitation of different animals.

Increases in extreme heat stress in domesticated livestock species during the twenty-first century.

Anthropogenic climate change is expected to have major impacts on domesticated livestock, including increased heat stress in animals in both intensive and extensive livestock systems. We estimate the changes in the number of extreme heat stress days per year for animals raised outdoors that can be expected in the major domesticated animal species (cattle, sheep, goats, poultry, and pigs) across the globe during this century. We used the temperature humidity index as a proxy for heat stress, calculated using temperature and relative humidity data collated from an ensemble of CMIP6 climate model output for mid and end century. We estimate changes in the proportions of different livestock species that may be at increased risk of extreme heat stress under two contrasting greenhouse gas emission scenarios. Results are discussed in relation to changes in the suitability of different climate conditions for domesticated livestock during the current century. We find that by end century, extreme heat stress risk is projected to increase for all livestock species in many parts of the tropics and some of the temperate zones, and to become climatically more widespread, compared to 2000. Although adaptation options exist for both intensive and extensive livestock production systems, the increasing pervasiveness of extreme heat stress risk in the future will seriously challenge the viability of outdoor livestock keeping, particularly in the lower latitudes in lower and middle‐income countries where the costs of adaptation may be challenging to address.

Domestic chicken diversity: Origin, distribution, and adaptation.

Chicken is the most numerous among the domesticated livestock species. Across cultures, religions, and societies, chicken is widely accepted with little or no taboo compared to other domestic animals. Its adaptability to diverse environmental conditions and demonstrated potential for breeding improvement provide a unique genetic resource for addressing the challenges of food security in a world impacted by climatic change and human population growth. Recent studies, shedding new knowledge on the chicken genomes, have helped reconstruct its past evolutionary history. Here, we review the literature concerning the origin, dispersion, and adaptation of domestic chicken. We highlight the role of human and natural selection in shaping the diversity of the species and provide a few examples of knowledge gaps that may be the focus of future research.

Controversies and Common Ground in Wild and Domestic Fine Fiber Production in Argentina

This work analyzes possible obstacles to developing new products or old merchandise using an innovative method. It will look into stakeholders of fine fiber and meat products from three distinctive socioecological systems. Through three case studies, we explore how natural resources management is connected to interests, values, and knowledge by stakeholders, which include government, the scientific community, and people with rural livelihoods. The government vertex is the national and provincial authorities involved with decision-makers at the national and provincial level. The Scientific-Technological vertex includes researchers from INTA, CONICET, and Universities. Rural livelihoods include livestock keepers, farmers, and local people with traditional knowledge. We will address the goods and services provided by two species of wild camelids and domestic livestock. The three cases have both similarities and differences in their focus and common ground of controversial spaces. They create complex networks of relationships and bonds leading to diverse outcomes. Top-down or bottom-up experiences hold distinct epistemology and research consequences, they affect rural livelihoods in various ways. For the three rural livelihoods, meaningful regulations should be endogenous social constructions. However, there are no longitudinal studies on the trajectories of these case studies. Long-term multispecies grazing opportunities are available for the three case studies. It depends on how stakeholders identify flexibility in their common ground to enable resilience to catastrophic events.

Reliable, scalable functional genetics in bloodstream-form Trypanosoma congolense in vitro and in vivo.

Animal African trypanosomiasis (AAT) is a severe, wasting disease of domestic livestock and diverse wildlife species. The disease in cattle kills millions of animals each year and inflicts a major economic cost on agriculture in sub-Saharan Africa. Cattle AAT is caused predominantly by the protozoan parasites Trypanosoma congolense and T. vivax, but laboratory research on the pathogenic stages of these organisms is severely inhibited by difficulties in making even minor genetic modifications. As a result, many of the important basic questions about the biology of these parasites cannot be addressed. Here we demonstrate that an in vitro culture of the T. congolense genomic reference strain can be modified directly in the bloodstream form reliably and at high efficiency. We describe a parental single marker line that expresses T. congolense-optimized T7 RNA polymerase and Tet repressor and show that minichromosome loci can be used as sites for stable, regulatable transgene expression with low background in non-induced cells. Using these tools, we describe organism-specific constructs for inducible RNA-interference (RNAi) and demonstrate knockdown of multiple essential and non-essential genes. We also show that a minichromosomal site can be exploited to create a stable bloodstream-form line that robustly provides >40,000 independent stable clones per transfection-enabling the production of high-complexity libraries of genome-scale. Finally, we show that modified forms of T. congolense are still infectious, create stable high-bioluminescence lines that can be used in models of AAT, and follow the course of infections in mice by in vivo imaging. These experiments establish a base set of tools to change T. congolense from a technically challenging organism to a routine model for functional genetics and allow us to begin to address some of the fundamental questions about the biology of this important parasite.

PSIII-14 Genetic Diversity and Haplogroup Distribution of Asian Chicken Populations using Mitochondrial DNA sequence

Abstract Estimating the genetic diversity and origins is critical for the identification and conservation of valuable animal genetic resources. Mitochondrial DNA (mtDNA) polymorphisms have played a significant role in tracing the origin of specific breeds and the genetic diversity of domestic chicken and other livestock species due to their maternal inheritance, greater rate of substitution relative to unclear genes, and much lower rate of recombination. In this study, we analyzed the mtDNA D-loop region of 457 genomic DNA samples from 28 Asian chicken populations (Korea: 6, Laos: 4, Bangladesh: 3, Cambodia: 3, Indonesia: 3, Vietnam: 2, Kyrgyzstan: 1, Mongolia: 1, Nepal: 1, Sri Lanka: 1 and White Leghorn, Cornish, Rhode Island Red) from ten Asian countries to clarify their genetic diversity and phylogenetic relationships. In addition, 49 reference data (43: Oka et al data, 6: red jungle fowl) were used. We found 82 haplotypes defined by 81 variable sites. Overall haplotype and nucleotide diversity were 0.978±0.01 and 0.0018, respectively. Phylogenetic analyses revealed seven haplogroups (Type A-G). The most of Asian chickens were included in haplogroup A(44.9%), B(22.8%), C(19.3%). Haplogroup F did not belong to any Asian chicken populations (include only 2 reference data). Mongolia chickens were only clustered in haplogroup A and Indonesian chickens did only haplogroup C. About half of Kyrgyzstani chicken populations (43.4%) were clustered in haplogroup E. Vietnamese chicken populations belong to all haplogroup except F. In case of red jungle fowl, G.g.gallus and G.g.bankiva were clustered in haplogroup C. G.g.spadiceus was clusterd in haplogroup E or A. As a result, it was possible to identify the mtDNA diversity and origin of Asian chicken populations. In future studies, it is necessary to understand the origin of each haplogroup by adding more reference data and red jungle fowl sequence data.

Convergent evolution of zoonotic Brucella species toward the selective use of the pentose phosphate pathway

Mechanistic understanding of the factors that govern host tropism remains incompletely understood for most pathogens. Brucella species, which are capable of infecting a wide range of hosts, offer a useful avenue to address this question. We hypothesized that metabolic fine-tuning to intrahost niches is likely an underappreciated axis underlying pathogens' ability to infect new hosts and tropism. In this work, we compared the central metabolism of seven Brucella species by stable isotopic labeling and genetics. We identified two functionally distinct groups, one overlapping with the classical zoonotic species of domestic livestock that exclusively use the pentose phosphate pathway (PPP) for hexose catabolism, whereas species from the second group use mostly the Entner-Doudoroff pathway (EDP). We demonstrated that the metabolic dichotomy among Brucellae emerged after the acquisition of two independent EDP-inactivating mutations in all classical zoonotic species. We then examined the pathogenicity of key metabolic mutants in mice and confirmed that this trait is tied to virulence. Altogether, our data are consistent with the hypothesis that the PPP has been incrementally selected over the EDP in parallel to Brucella adaptation to domestic livestock.

Virulence beneath the fleece; a tale of foot-and-mouth disease virus pathogenesis in sheep

Foot-and-mouth disease virus (FMDV) is capable of infecting all cloven-hoofed domestic livestock species, including cattle, pigs, goats, and sheep. However, in contrast to cattle and pigs, the pathogenesis of FMDV in small ruminants has been incompletely elucidated. The objective of the current investigation was to characterize tissue- and cellular tropism of early and late stages of FMDV infection in sheep following three different routes of simulated natural virus exposure. Extensive post-mortem harvest of tissue samples at pre-determined time points during early infection (24 and 48 hours post infection) demonstrated that tissues specifically susceptible to primary FMDV infection included the paraepiglottic- and palatine tonsils, as well as the nasopharyngeal mucosa. Additionally, experimental aerosol inoculation of sheep led to substantial virus replication in the lungs at 24-48 hours post-inoculation. During persistent infection (35 days post infection), the paraepiglottic- and palatine tonsils were the only tissues from which infectious FMDV was recovered. This is strikingly different from cattle, in which persistent FMDV infection has consistently been located to the nasopharyngeal mucosa. Analysis of tissue sections by immunomicroscopy revealed a strict epithelial tropism during both early and late phases of infection as FMDV was consistently localized to cytokeratin-expressing epithelial cells. This study expands upon previous knowledge of FMDV pathogenesis in sheep by providing detailed information on the temporo-anatomic distribution of FMDV in ovine tissues. Findings are discussed in relation to similar investigations previously performed in cattle and pigs, highlighting similarities and differences in FMDV pathogenesis across natural host species.

Systematic reviews and meta-analyses in animal health, performance, and on-farm food safety: a scoping review.

Systematic reviews and meta-analyses are used to summarize and interpret evidence for clinical decision-making in human health. The extent of the application of these methods in veterinary medicine and animal agriculture is unknown. The goal of this scoping study was to ascertain trends in the publication of systematic reviews and meta-analyses examining animal health, animal performance, and on-farm food safety. Online databases were searched for reviews published between 1993 and 2018 that focused on relevant outcomes in domestic livestock, companion animals, or wildlife species. In total 1787 titles and abstracts underwent data characterization. Dairy cattle, fish, and pigs were the most common target commodity groups. Few articles investigated both health and performance outcomes (only health: n = 418; only performance: n = 701; both health and performance: n = 103). Most of the reviews (67.6%, n = 1208/1787) described a meta-analysis but did not state in the title or abstract that a systematic review was also conducted, which is potentially problematic. Adherence to reporting guidelines is recommended for all systematic reviews and meta-analyses. For research areas with many reviews, an evidence repository is recommended. For less well-reviewed areas, additional investigation may be necessary to identify the reasons for the lack of synthesis research.

Impact of Ergot Alkaloids on Female Reproduction in Domestic Livestock Species.

Fescue toxicosis is a multifaceted syndrome that elicits many negative effects on livestock consuming ergot alkaloids produced by endophyte-infected tall fescue. The economic losses associated with fescue toxicosis are primarily due to reproductive failure including altered cyclicity, suppressed hormone secretion, reduced pregnancy rates, agalactia, and reduced offspring birth weights. For decades, a multitude of research has investigated the physiological and cellular mechanisms of these reproductive failures associated with fescue toxicosis. This review will summarize the various effects of ergot alkaloids on female reproduction in grazing livestock species.

The sero-epidemiology of Coxiella burnetii (Q fever) across livestock species and herding contexts in Laikipia County, Kenya.

Coxiella burnetii, the causative agent of Query fever (Q fever), is among the most highly infectious zoonotic pathogens transmitted among livestock, with chronic effects challenging to veterinary and medical detection and care systems. Transmission among domestic livestock species can vary regionally due to herd management practices that determine which livestock species are raised, whether or not livestock are in contact with wildlife, and the susceptibility of these livestock to infection. To explore how different livestock management practices are associated with the risk of infection in multispecies environments, we carried out a comparative study of three types of herd management systems in the central Kenyan county of Laikipia: agro‐commercial, mixed conservancy/commercial, and smallholder ranches. We tested C. burnetii antibody seroprevalence in four common livestock species. Across all management types, the highest seroprevalence was in camels (20%), followed by goats (18%), sheep (13%), and cattle (6%). We observed a lower odds of testing seropositive for young compared to adult animals (adjusted OR = 0.44 [95% CI 0.24, 0.76]), and for males compared to females (adjusted OR = 0.52 [95% CI 0.33, 0.80]). Animals from mixed conservancy/commercial and smallholder operations had a higher odds of testing seropositive compared to animals from agro‐commercial ranches (adjusted OR = 5.17 [95% CI 2.71, 10.44] and adjusted OR = 2.21 [95% CI 1.17, 4.43] respectively). These data suggest that herd management practices might affect the transmission dynamics of C. burnetiiin arid African ecosystems like those seen in Kenya where several transmission modes are possible, risk of drought has promoted new livestock species such as camels, and multiple wildlife species may co‐occur with livestock on the landscape. Further longitudinal studies are needed to disentangle the mechanisms underlying these patterns, and further explore transmission patterns between wildlife, domestic animal, and human populations.

Potential Intermediate Hosts for Coronavirus Transmission: No Evidence of Clade 2c Coronaviruses in Domestic Livestock from Ghana.

The emergence of Middle East Respiratory Syndrome Coronavirus (MERS-CoV), nearly a decade ago with worldwide distribution, was believed to be of zoonotic origin from bats with dromedary camels as intermediate hosts. There is a likelihood of other domestic livestock serving as intermediate hosts for this virus. The presence of coronaviruses, closely related to MERS-CoV in Ghanaian bats, presented the opportunity to test the hypothesis of transmissibility of this virus through domestic livestock species. The possible interactions between livestock and bats in 31 household farms were accessed by observation and interviews with farmers. Rectal swabs and serum from cattle, sheep, goats, donkeys, and swine from commercial and household farms were tested for MERS-CoV and a Nycteris sp. bat coronavirus, previously detected in Ghana. A pan-PCR assay to detect clade 2c viruses and recombinant immunofluorescence assay to detect anti-spike IgG antibodies against the target viruses were used. Likely contact between livestock and bats was determined for 13 farms (41.9%) that reported confining their livestock and also observing bats in their homes. Livestock were left unconfined on eight farms (25.8%) that also observed bats roosting in trees close to their homes. No viral RNA or antibodies against the two coronaviruses were detected in any of the livestock species tested. Cattle, sheep, goats, donkeys, and swine are not likely hosts of clade 2c coronaviruses.

Development of a Whole-Virus ELISA for Serological Evaluation of Domestic Livestock as Possible Hosts of Human Coronavirus NL63.

Known human coronaviruses are believed to have originated in animals and made use of intermediate hosts for transmission to humans. The intermediate hosts of most of the human coronaviruses are known, but not for HCoV-NL63. This study aims to assess the possible role of some major domestic livestock species as intermediate hosts of HCoV-NL63. We developed a testing algorithm for high throughput screening of livestock sera with ELISA and confirmation with recombinant immunofluorescence assay testing for antibodies against HCoV-NL63 in livestock. Optimization of the ELISA showed a capability of the assay to significantly distinguish HCoV-NL63 from HCoV-229E (U = 27.50, p < 0.001) and HCoV-OC43 (U = 55.50, p < 0.001) in coronavirus-characterized sera. Evaluation of the assay with collected human samples showed no significant difference in mean optical density values of immunofluorescence-classified HCoV-NL63-positive and HCoV-NL63-negative samples (F (1, 215) = 0.437, p = 0.509). All the top 5% (n = 8) most reactive human samples tested by ELISA were HCoV-NL63 positive by immunofluorescence testing. In comparison, only a proportion (84%, n = 42) of the top 25% were positive by immunofluorescence testing, indicating an increased probability of the highly ELISA reactive samples testing positive by the immunofluorescence assay. None of the top 5% most ELISA reactive livestock samples were positive for HCoV-NL63-related viruses by immunofluorescence confirmation. Ghanaian domestic livestock are not likely intermediate hosts of HCoV-NL63-related coronaviruses.

Lack of Transmission of Foot-and-Mouth Disease Virus From Persistently Infected Cattle to Naïve Cattle Under Field Conditions in Vietnam.

Foot-and-mouth disease (FMD), caused by FMD virus (FMDV; Aphthovirus, Picornaviridae), is a highly contagious and economically important disease of cloven-hoofed domestic livestock and wildlife species worldwide. Subsequent to the clinical phase of FMD, a large proportion of FMDV-infected ruminants become persistently infected carriers, defined by detection of FMDV in oropharyngeal fluid (OPF) samples 28 days or more post-infection. The goal of this prospective study was to characterize the FMD carrier state in cattle subsequent to natural infection under typical husbandry practices in Vietnam. Ten persistently infected cattle on eight farms in the Long An province in southern Vietnam were monitored by monthly screening of serum and oropharyngeal fluid samples for 12 months. To assess transmission from FMDV carriers, 16 naïve cattle were intentionally brought into direct contact with the persistently infected animals for 6 months, and were monitored by clinical and laboratory methods. The restricted mean duration of the FMD carrier state was 27.7 months, and the rate of decrease of the proportion of carrier animals was 0.03 per month. There was no evidence of transmission to naïve animals throughout the study period. Additionally, there was no detection of FMDV infection or seroconversion in three calves born to carrier animals during the study. The force of infection for carrier-to-contact transmission was 0 per month, with upper 95% confidence limit of 0.064 per month. Phylogenetic analysis of viral protein 1 (VP1) coding sequences obtained from carriers indicated that all viruses recovered in this study belonged to the O/ME-SA/PanAsia lineage, and grouped phylogenetically with temporally and geographically related viruses. Analysis of within-host evolution of FMDV, based upon full-length open reading frame sequences recovered from consecutive samples from one animal, indicated that most of the non-synonymous changes occurred in Lpro, VP2, and VP3 protein coding regions. This study suggests that the duration of FMDV persistent infection in cattle may be longer than previously recognized, but the risk of transmission is low. Additional novel insights are provided into within-host viral evolution under natural conditions in an endemic setting.

Genetic structure in Red Junglefowl (Gallus gallus) populations: Strong spatial patterns in the wild ancestors of domestic chickens in a core distribution range.

Red Junglefowl (Gallus gallus) are among the few remaining ancestors of an extant domesticated livestock species, the domestic chicken, that still occur in the wild. Little is known about genetic diversity, population structure, and demography of wild Red Junglefowl in their natural habitats. Extinction threats from habitat loss or genetic alteration from domestic introgression exacerbate further the conservation status of this progenitor species. In a previous study, we reported extraordinary adaptive genetic variation in the MHC B‐locus in wild Red Junglefowl and no evidence of allelic introgression between wild and domestic chickens was observed. In this study, we characterized spatial genetic variation and population structure in naturally occurring populations of Red Junglefowl in their core distribution range in South Central Vietnam. A sample of 212 Red Junglefowl was obtained from geographically and ecologically diverse habitats across an area of 250 × 350 km. We used amplified fragment‐length polymorphism markers obtained from 431 loci to determine whether genetic diversity and population structure varies. We found that Red Junglefowl are widely distributed but form small and isolated populations. Strong spatial genetic patterns occur at both local and regional scales. At local scale, population stratification can be identified to approximately 5 km. At regional scale, we identified distinct populations of Red Junglefowl in the southern lowlands, northern highlands, and eastern coastal portions of the study area. Both local and long‐distance genetic patterns observed in wild Red Junglefowl may reflect the species’ ground‐dwelling and territorial characteristics, including dispersal barriers imposed by the Annamite Mountain Range. Spatially explicit analyses with neutral genetic markers can be highly informative and here elevates the conservation profile of the wild ancestors of domesticated chickens.

Evaluation of ‘recombinant secretary antigens’ based ‘cocktail ELISA’ for the diagnosis of Johne's disease and to differentiate non-infected, infected and vaccinated goats in combination with indigenous ELISA test

Johne's disease (JD) is the most widely distributed disease of the domestic livestock species world-wide. In the absence of control programs, bio-load of Mycobacterium aviumsubspecies paratuberculosis (MAP) had shown increasing trends in the Indian goat population over the years. Recently a therapeutic 'Indigenous vaccine' has been developed against JD in goat herds. Before initiating wide-scale use of 'indigenous vaccine' in the field as part of National Johne's disease control program, it is essential to have a test, which can differentiate non-infected, infected and vaccinated goats. In this study, a new 'cocktail ELISA' using six specific recombinant secretary proteins (RSPs) or culture filtrate proteins (CFPs) (MAP 1693c, MAP 2168c, MAP Mod D, MAP 85c, MAP Pep AN and MAP Pep AC) was developed first time in India. The RSPs based 'cocktail ELISA' was evaluated for the diagnosis of JD and c_ELISA along with i_ELISA, successfully differentiated non-infected, infected and vaccinated goats, thus facilitating the use of JD vaccine in the field for the control of the MAP infection in the goat population of the country.

Comparison of fecal egg counting methods in four livestock species

Gastrointestinal nematode parasites are important pathogens of all domesticated livestock species. Fecal egg counts (FEC) are routinely used for evaluating anthelmintic efficacy and for making targeted anthelmintic treatment decisions. Numerous FEC techniques exist and vary in precision and accuracy. These performance characteristics are especially important when performing fecal egg count reduction tests (FECRT). The objective of this study was to compare the accuracy and precision of three commonly used FEC methods and determine if differences existed among livestock species. In this study, we evaluated the modified-Wisconsin, 3-chamber (high-sensitivity) McMaster, and Mini-FLOTAC methods in cattle, sheep, horses, and llamas in three phases. In the first phase, we performed an egg-spiking study to assess the egg recovery rate and accuracy of the different FEC methods. In the second phase, we examined clinical samples from four different livestock species and completed multiple replicate FEC using each method. In the last phase, we assessed the cheesecloth straining step as a potential source of egg loss.In the egg-spiking study, the Mini-FLOTAC recovered 70.9% of the eggs, which was significantly higher than either the McMaster (P = 0.002) or Wisconsin (P = 0.002). In the clinical samples from ruminants, Mini-FLOTAC consistently yielded the highest EPG, revealing a significantly higher level of egg recovery (P < 0.0001). For horses and llamas, both McMaster and Mini-FLOTAC yielded significantly higher EPG than Wisconsin (P < 0.0001, P < 0.0001, P < 0.001, and P = 0.024). Mini-FLOTAC was the most accurate method and was the most precise test for both species of ruminants. The Wisconsin method was the most precise for horses and McMaster was more precise for llama samples. We compared the Wisconsin and Mini-FLOTAC methods using a modified technique where both methods were performed using either the Mini-FLOTAC sieve or cheesecloth. The differences in the estimated mean EPG on log scale between the Wisconsin and mini-FLOTAC methods when cheesecloth was used (P < 0.0001) and when cheesecloth was excluded (P < 0.0001) were significant, providing strong evidence that the straining step is an important source of error. The high accuracy and precision demonstrated in this study for the Mini-FLOTAC, suggest that this method can be recommended for routine use in all host species. The benefits of Mini-FLOTAC will be especially relevant when high accuracy is important, such as when performing FECRT.