Advancements in Understanding and Enhancing Antioxidant-Mediated Sperm Cryopreservation in Small Ruminants: Challenges and Perspectives.

Cryopreservation poses significant challenges to the preservation of sperm integrity and function, particularly in small ruminants where cryodamage is pronounced. This review explores the molecular mechanisms underlying sperm cryodamage and strategies for improving cryopreservation outcomes, with a focus on the role of antioxidants. Cryopreservation-induced alterations in proteins and RNA transcripts critical for sperm function, including motility, capacitation, fertilization, and embryo development, are discussed. Proteomic, transcriptomic, and epigenomic advancements have provided valuable insights into these mechanisms, offering potential biomarkers for predicting sperm freezability and enhancing cryopreservation strategies. Combining technologies such as mass spectrometry and flow cytometry allows for a comprehensive understanding of molecular and cellular changes induced by the freezing-thawing process. However, challenges remain in optimizing cryoprotectant formulations and antioxidant supplementation to improve post-thaw sperm fertility. Further research is needed to explore a wider range of novel cryoprotectants, antioxidants, and proteins for cryopreservation media, as well as to validate their efficacy in enhancing sperm viability and function. Additionally, investigations into the effects of cryopreservation on RNA transcripts and epigenetic factors in small ruminant species are warranted to advance our understanding of sperm preservation. Overall, this review highlights the importance of antioxidants in mitigating cryodamage and underscores the need for continued research to refine cryopreservation protocols and improve reproductive outcomes in small ruminants.

Prevalence of Helminths in Small Ruminant Farms and Evaluation of Control Practices Used to Counter Anthelmintic Resistance in Southern Italy.

Anthelmintic resistance in small ruminants is a serious worldwide problem. To reduce their spread, it is essential to know the prevalence of helminths on farms and the control practices adopted. As these studies in the Calabria region of southern Italy are fragmentary and outdated, a study on the prevalence of helminths in small ruminant holdings in this area has been conducted. The measures implemented to control helminths were also evaluated through questionnaires administered to farmers. In particular, on 90 farms (45 sheep and 45 goats), 1800 faecal samples from 900 sheep and 900 goats were collected in the spring. Using the FLOTAC dual technique, parasitological examinations demonstrated the presence of gastrointestinal nematodes in 100% of sheep and goat farms, followed by Nematodirus spp. (84.44% sheep and 48.89% goats), Moniezia spp. (73.33% sheep and 35.56% goats), Trichuris ovis (48.89% sheep and 42.22% goats), lungworms (28.89% sheep and 42.22% goats), Strongyloides papillosus (40% sheep and 26.67% goats), Dicrocoelium dendriticum (13.33% sheep and 26.67% goats), Calicophoron daubneyi (6.67% sheep and 31.11% goats), Fasciola hepatica (6.67% sheep and 4.44% goats), and Skrjabinema ovis (4.44% sheep and goats). The questionnaires showed that 82% and 85% of the farmers had applied pasture rotation, and that 93.3% and 86.6% had used anthelmintics in the previous year for sheep and goats, respectively. Only 24.4% of sheep farmers and 11.3% of goat farmers had carried out parasitological tests prior to treatments. The most used classes of anthelmintics were macrocyclic lactones and benzimidazoles, and only in 21.6% and 15.6%, for sheep and goats, respectively, was drug rotation carried out. These results denote that helminths represent a health problem for small ruminants and highlight a lack of knowledge of parasite control strategies among farmers. In these conditions, anthelmintic resistance phenomena could develop over time. Therefore, it is necessary to implement all possible strategies for the control of helminths, and to prevent the spread of anthelmintic resistance phenomena on farms in southern Italy.

Recapitulation of Peste des Petits Ruminants (PPR) Prevalence in Small Ruminant Populations of Pakistan from 2004 to 2023: A Systematic Review and Meta-Analysis.

Peste des petits ruminants (PPR) is an extremely transmissible viral disease caused by the PPR virus that impacts domestic small ruminants, namely sheep and goats. This study aimed to employ a methodical approach to evaluate the regional occurrence of PPR in small ruminants in Pakistan and the contributing factors that influence its prevalence. A thorough search was performed in various databases to identify published research articles between January 2004 and August 2023 on PPR in small ruminants in Pakistan. Articles were chosen based on specific inclusion and exclusion criteria. A total of 25 articles were selected from 1275 studies gathered from different databases. The overall pooled prevalence in Pakistan was calculated to be 51% (95% CI: 42-60), with heterogeneity I2 = 100%, τ2 = 0.0495, and p = 0. The data were summarized based on the division into five regions: Punjab, Baluchistan, KPK, Sindh, and GB and AJK. Among these, the pooled prevalence of PPR in Sindh was 61% (95% CI: 46-75), I2 = 100%, τ2 = 0.0485, and p = 0, while in KPK, it was 44% (95% CI: 26-63), I2 = 99%, τ2 = 0.0506, and p < 0.01. However, the prevalence of PPR in Baluchistan and Punjab was almost the same. Raising awareness, proper surveillance, and application of appropriate quarantine measures interprovincially and across borders must be maintained to contain the disease.

Inseminate without hormonal treatment in dairy sheep farms: exploring the consequences on the sustainability of several contrasted production systems.

Alternatives to hormonal treatments (HTs) in dairy sheep reproduction management are being explored in response to increasing societal concerns regarding animal welfare and food safety. However, hormone-free reproduction implies higher variability in flock performances and additional constraints for timely synchronised artificial insemination (AI) in the flock, impacting the diffusion of genetic progress. The use of the male effect, a well-known practice to induce synchronised oestrus, combined with precision tools (e.g., heat detector), is a plausible way to implement AI without HT in dairy sheep farms. To date, the consequences of such alternative reproduction management on the whole farm sustainability remain unknown. To anticipate these potential impacts, a multiagent model (REPRIN'OV) was used to simulate dairy sheep farms' sustainability indicators (biotechnical, economic, environmental and workload). A reproduction management scenario, including the use of the male effect followed by AI on the adult ewes (HFAI), was simulated and compared to the current reproduction management of four case study farms (Early_conv, Late_conv, Early_org and Late_org). They were selected to represent the different agricultural models (Conventional or Organic) and reproduction seasons (Early - during spring, out of ewes' natural reproduction season - or Late -from early summer to the end of autumn) of the Roquefort Basin's farms in Southern France. Simulation results showed that the HFAI scenario had different consequences depending on the farm's production system type. A negative effect on most key sustainability indicators of the Conv farms was observed, as a significant reduction in the fertility rate, in the proportion of young ewes born from AI (-54% in both farms; P < 0.05) and in the flock's milk production were observed; while the workload and greenhouse gas (GHG) emissions were increased compared to the initial scenario. In the Org farms, HFAI had neutral to positive effects on most indicators as the fertility, milk production of the flock, workload during milking and GHG emissions were barely affected by this scenario, while an increase in the proportion of young ewes born from AI was observed (+39% and + 43% in each farm, respectively; P < 0.05), allowing a better farm gross margin. Still, the workload during lambing was increased in Early_org (+18%; P < 0.05), as Early farms, tended to be more negatively impacted by HFAI than Late ones. Overall, our simulation approach provides interesting elements to exchange with stakeholders on how to progress towards a socially acceptable reproduction management system, for the dairy sheep sector.

Prevalence of Gastrointestinal Parasites in Small Ruminant Farms in Southern Spain.

The primary population of small ruminants in Spain is concentrated in the southern region, a critical area for the country's livestock production. Indirect economic losses can occur when this livestock is affected by gastrointestinal parasites. This study aimed to determine the prevalence of these parasites in small ruminant herds (159 sheep and 39 goats) through coprological analyses and conducted a survey on farmers' management practices related to gastrointestinal parasite control. The survey results revealed some important aspects: monitoring through coprological analyses is not a common practice; veterinarians are not typically involved in deworming plans; anthelmintic treatment in adults is often applied twice a year in sheep and once a year in goats; and finally, drug rotation was higher in sheep farms. Coprological analyses showed Eimeria spp. as the most common parasitic infection, followed by Strongyles infection. Other parasites like Moniezia spp., Trichuris spp., and D. dendriticum were less important, although their prevalence was higher in sheep than goats. This constitutes the first report on the epidemiological status of gastrointestinal parasites in small ruminants in southern Spain. Based on the survey findings, the introduction of certain management measures on farms could potentially mitigate parasite infections.

Assessment of health problems of sheep and goats based on ante-mortem and post-mortem inspection at Addis Ababa Abattoir, Ethiopia.

Introduction: Ethiopia has a rapidly growing small ruminant sector, which faces low productivity due to husbandry practices and poor health condition of the animals. A study was conducted in Ethiopia's largest municipal abattoir with the objective to assess the health problems of sheep and goats presented for slaughter using standard ante-mortem and post-mortem methodology. Methods: A cross-sectional study using systematic random sampling was conducted on 384 sheep and 384 goats from January to July 2014. Results: Soiled skin (69.1%), poor body condition (24.3%), and nostril discharge (19.5%) were common among both species at ante-mortem examination. Gross lesions were frequent in livers (39.7%) and lungs (37.2%), while pneumonia (18.1%) and adhesions (13.8%) were frequent in the lungs of sheep and goats, indicating stress-related illness. Parasitic lesions, especially fasciolosis (19.3%) and hydatid cysts (8.1%) were significantly more common in sheep livers (p 0.05). The direct financial loss from lesions in both species was 1,077,015 ETB or 53,851 USD per year, most of which was estimated to occur from carcass bruising. Discussion: The findings indicate that reducing parasite burden and preventing carcass bruising through improved handling could significantly increase the profitability of the small ruminant meat sector in Ethiopia.

Molecular Epidemiology and Phylogeny of Theileria ovis and Theileria lestoquardi in Sheep and Goats from Southern Punjab, Pakistan.

Background: Theileria spp. are responsible for ovine and caprine theileriosis, leading to significant morbidity and mortality in small ruminants. The present study aims to investigate Theileria spp. infections in small ruminants from Southern Punjab in Pakistan, and genetic characterize revealed Theileria spp. isolates. Methods: A total of 93 sheep and 107 goats were sampled between May and August 2022. Blood smears were examined microscopically, and PCR amplification targeting the 18S rRNA gene was performed to detect Theileria spp. Additionally, specific PCR assays targeting 18S rRNA and ms1 partial sequences were used to identify Theileria ovis and T. lestoquardi, respectively.  Results: The prevalence of Theileria spp. was significantly higher using PCR (13.5%) compared to microscopic screening (5%). Sheep showed a higher prevalence rate (19.4%) compared to goats (8.4%) (p = 0.024). Young sheep aged ≤ 1 year were more commonly infected with Theileria spp. (41%) compared to older sheep (p = 0.006). The prevalence of Theileria spp. was higher in sheep-only herds (37.3%) compared to goat-only herds (18%) or mixed-species herds (8.1%) (p = 0.015). The prevalence rates of T. ovis and T. lestoquardi were 9% and 2.5%, respectively, with four animals (2 goats and 2 sheep) showing co-infection. Phylogenetic analysis revealed that our T. ovis 18S rRNA sequence clustered with previously reported sequences from sheep in Turkey, China, Spain, and goats in Tanzania. The obtained T. lestoquardi ms1 partial sequence formed a distinct cluster from other T. lestoquardi isolates in Pakistan and neighboring countries.  Conclusion: Theileria spp. co-circulation in Pakistani small ruminants, particularly the presence of T. ovis and T. lestoquardi, highlights the need for attention from animal health decision-makers due to their financial and health impacts.

Ovine and Caprine Strains of Corynebacterium pseudotuberculosis on Czech Farms-A Comparative Study.

Caseous lymphadenitis (CLA) is a worldwide disease of small ruminants caused by Corynebacterium pseudotuberculosis, a facultative intracellular pathogen that is able to survive and multiply in certain white blood cells of the host. In this study, 33 strains of C. pseudotuberculosis were isolated from sheep and goats suffering from CLA on nine farms in the Czech Republic. All these strains were tested for their antibiotic susceptibility, ability to form a biofilm and resistance to the effects of commonly used disinfectant agents. To better understand the virulence of C. pseudotuberculosis, the genomes of strains were sequenced and comparative genomic analysis was performed with another 123 genomes of the same species, including ovis and equi biovars, downloaded from the NCBI. The genetic determinants for the virulence factors responsible for adherence and virulence factors specialized for iron uptake and exotoxin phospholipase D were revealed in every analyzed genome. Carbohydrate-Active Enzymes were compared, revealing the presence of genetic determinants encoding exo-α-sialidase (GH33) and the CP40 protein in most of the analyzed genomes. Thirty-three Czech strains of C. pseudotuberculosis were identified as the biovar ovis on the basis of comparative genome analysis. All the compared genomes of the biovar ovis strains were highly similar regardless of their country of origin or host, reflecting their clonal behavior.

Multi-omics reveal the gut microbiota-mediated severe foraging environment adaption of small wild ruminants in the Three-River-Source National Park, China.

The Tibetan antelope (Pantholops hodgsonii), blue sheep (Pseudois nayaur), and Tibetan sheep (Ovis aries) are the dominant small ruminants in the Three-River-Source National Park (TRSNP). However, knowledge about the association between gut microbiota and host adaptability remains poorly understood. Herein, multi-omics sequencing approaches were employed to investigate the gut microbiota-mediated forage adaption in these ruminants. The results revealed that although wild ruminants (WR) of P. hodgsoni and P. nayaur were faced with severe foraging environments with significantly low vegetation coverage and nutrition, the apparent forage digestibility of dry matter, crude protein, and acid detergent fiber was significantly higher than that of O. aries. The 16s rRNA sequencing showed that the gut microbiota in WR underwent convergent evolution, and alpha diversity in these two groups was significantly higher than that in O. aries. Moreover, indicator species, including Bacteroidetes and Firmicutes, exhibited positive relationships with apparent forage digestibility, and their relative abundances were enriched in the gut of WR. Enterotype analysis further revealed that enterotype 1 belonged to WR, and the abundance of fatty acid synthesis metabolic pathway-related enzyme genes was significantly higher than enterotype 2, represented by O. aries. Besides, the metagenomic analysis identified 14 pathogenic bacterial species, among which 10 potentially pathogenic bacteria were significantly enriched in the gut microbiota of O. aries. Furthermore, the cellulolytic strains and genes encoding cellulase and hemicellulase were significantly enriched in WR. In conclusion, our results provide new evidence of gut microbiota to facilitate wildlife adaption in severe foraging environments of the TRSNP, China.

Solanum glaucophyllum intoxication in goats: Case study.

Solanum glaucophyllum is a toxic plant with calcinogenic effect that causes enzootic calcinosis (EC) characterized by soft tissue metastatic mineralization mainly in cattle and rarely sheep, buffaloes, pigs, horses, and goats. We describe an outbreak of EC in a herd of 64 goats due to S. glaucophyllum consumption. Thirty-four goats were affected exhibiting hirsutism, stiffening, kyphosis and emaciation. Twelve goats died. Grossly, tissue mineralization was observed in the aorta and carotid arteries, lungs, and heart. Lesions were characterized by multiple rough white plaques, and hardened tissues with loss of elasticity. Microscopically, multisystemic mineralization was observed in aorta and carotid arteries, heart, lung, abomasum, intestine, spleen, lymph nodes, kidney, spleen, and meninges, characterized by extensive granular basophilic deposits of tunica media and/or intima of blood vessels; confirmed as calcium salt deposits with Von Kossa stain. We conclude that ingestion of S. glaucophyllum can cause EC in goats. Though EC is rare in goats under some conditions such as heavy drought and abundant S. glaucophyllum exposure disease can develop.

Ectoparasites Infestation to Small Ruminants and Practical Attitudes among Farmers toward Acaricides Treatment in Central Region of Java, Indonesia.

Ectoparasite infestations are one of the major problems affecting goat and sheep farming. Disease resulting from these infestations can cause changes in physical appearance, such as severe lesions on the skin, and economic consequences in the form of significantly reduced selling prices. This study aimed to determine the prevalence of ectoparasites in the Boyolali district, Central Java, Indonesia. A total of 651 sheep and goats were surveyed in this study. The parasites were collected via skin scraping, twister, or manually from clinically infected goats and sheep in traditional farms. All of the ectoparasites collected were successfully identified. The prevalence of ectoparasites in ruminants in Boyolali was 97.8% (637/651). The species make-up was as follows: Bovicola caprae 97.8% (637/651), Linognathus africanus 39% (254/651), Haemaphysalis bispinosa 3.5% (23/651), Ctenocephalides spp. 0.2% (1/651), and Sarcoptes scabiei 5.2% (34/651). The predilection sites were in the face, ear, and leg areas, and in the axillary, dorsal, abdomen, and scrotum regions of the surveyed animals. An evaluation of farmers' attitudes to ectoparasites was performed using a questionnaire. The findings of this study imply that animals in the investigated area are highly exposed to ectoparasite infestations. Given the importance of ectoparasites in both livestock and human communities, specifically in the health domain, more research into appropriate control strategies is necessary.

Global and regional seroprevalence of coxiellosis in small ruminants: A systematic review and meta-analysis.

BACKGROUND: Coxiellosis is a neglected zoonosis for occupationally exposed people in many parts of the world. Sheep and goats are two important small ruminants that act as reservoirs for human contamination; however, there is a lack of comprehensive data on the epidemiological aspects of coxiellosis in sheep and goats at regional and global levels. The aim of this study was to systematically review the available articles on seroprevalence of coxiellosis in sheep and goats and estimate the overall seroprevalence in different regions. METHODS: A systematic search strategy was performed in five electronic repositories for articles published until December 2021. Relevant data were extracted from the selected articles based on the inclusion criteria. A random effect meta-analysis model was used to analyse the data. Results are presented as the prevalence of seropositivity as a percentage and 95% confidence intervals. RESULTS: The global pooled seroprevalence of coxiellosis in sheep was 17.38% (95% confidence interval [CI]: 15.59%-19.17%). Overall, the regional level pooled prevalence estimates in sheep ranged from 15.04% (95% CI: 7.68%-22.40%) to 19.14% (95% CI: 15.51%-22.77%), depending on region. The global pooled seroprevalence of coxiellosis in goats was 22.60% (95% CI: 19.54%-25.66%). Overall, the regional level pooled prevalence estimates in goats ranged from 6.33% (95% CI: 2.96%-9.71%) to 55.13% (95% CI: 49.61%-60.65%), depending on the region. The prevalence estimates also varied significantly in both sheep and goats depending on age, sex, and rearing systems of the animals (p < 0.001). CONCLUSION: Seroprevalence of coxiellosis in both sheep and goats is considerable. Routine monitoring of the sheep and goat populations is needed to prevent spillover infection in other livestock and humans.

Seroepidemiology and associated risk factors of brucellosis in small ruminants of district Khanewal, Pakistan.

Objectives: Keeping in view the economic and veterinary public health importance of brucellosis, this research was conducted to determine its seroprevalence and associated risk determinants in small ruminants in district Khanewal, Southern Punjab, Pakistan. Materials and Methods: Two-stage cluster sampling technique was used for sampling, and the sample size was calculated using C-survey 2.0. Accordingly, sera samples (n = 392) were collected from small ruminants in the study area from October 2022 to July 2023. All the samples were tested for the presence of anti-Brucella antibodies by Rose Bengal Plate Test (RBPT), followed by confirmation of all the samples using an enzyme linked immunosorbent assay (ELISA) kit (ID.vet®, France; sensitivity and specificity=100%, each). Results: The seropositivity rate of brucellosis was 7.14% [n = 28/392; 95% confidence interval (CI) = 4.87%-10.12%] by RBPT, whereas the results of ELISA showed an overall seroprevalence rate of 7.40% (n = 29/392; 95% CI = 5.11%-10.37%) in the study population. Univariate analysis of risk factors revealed that abortion history (AH), retained fetal membranes (RFMs), repeat breeding, flock size (FS), educational status of farmers (ESFs), awareness about brucellosis (AB), and farm hygiene had a significant association with the seroprevalence of brucellosis (p < 0.05). The multivariate analysis using a binary logistic regression model revealed that variables including tehsil, FS, AH, RFM, ESF, AB, and farming system were significant factors (p < 0.05) associated with brucellosis in the target population. Conclusion: Brucellosis is prevalent in small ruminants in Khanewal, Pakistan. The disease burden can be reduced by improving the reproductive health of animals, farm hygiene, and farmers' awareness about the diseases. Further studies are needed on a larger scale to devise stringent disease control strategies to avoid losses associated with brucellosis at regional, national, and global levels.

Gastrointestinal anthelmintic plants used on small ruminants in Benin: Traditional use and scientific results - Review.

In Benin, livestock breeders frequently use medicinal plants to treat gastrointestinal diseases in small ruminants. The aim of this review is to list the plants traditionally used in this context and to present the scientific findings on the efficacy of these plants. An extensive search was carried out using PubMed, Scopus, ScienceDirect, Biomed Central and Google Scholar databases to collect data, with combinations of relevant french and english keywords such as "ethnobotanical survey", "anthelmintic properties", "medicinal plants", "gastrointestinal parasites", "digestive strongyles", "Haemonchus", "Trichostrongylus", "small ruminants", "sheep", "goats" and "Benin". A total of 45 published articles met the eligibility criteria. This review listed 123 plants used by breeders to treat gastrointestinal ailments in small ruminants. The most commonly used parts are leaves and barks, and the most common forms are decoction, maceration and powder. Scientific studies have demonstrated the anthelmintic properties of 18 plants, including Zanthoxylum zanthoxyloides, Newbouldia laevis, Mitragyna inermis and Combretum glutinosum. The powders or leaf extracts of these plants showed in vivo significant reductions of over 50% in egg excretion, larval establishment, viability and fertility of gastrointestinal strongyles in small ruminants. Extracts of these plants also revealed in vitro inhibitory activity of over 50% on egg hatching, larval migration and motility of gastrointestinal strongyles. This manuscript highlights the traditional use of anthelmintic plants in small ruminants in Benin and provides scientific results supporting the efficacy of these plants.

Synergistic Interaction Effect of Artemisia cina n-hexane Extract and Tagetes lucida Ethyl Acetate Extract on Haemonchus Contortus.

PURPOSE: We analysed the possible synergistic activity among active extracts from Artemisia cina and Tagetes lucida combinations on Haemonchus contortus, a nematode parasitising sheep. METHODS: The work was carried out in vitro on eggs and infective larvae (L3) of H. contortus. The results were analysed with SAS 9.1, applying the ANOVA and Tukey test, and the lethal concentration (LC) values LC50 and LC90 were determined with regression analysis, employing Proc Probit of SAS 9.1. Additionally, the lethal concentration (LC) was calculated with LC50 and LC90 to determine the synergistic effect. RESULTS: The results demonstrated a high efficacy of the two plants studied on both nematode eggs and L3 larvae as well as of their combinations. The highest egg hatching inhibition was obtained with a 50/50 combination, and the best larvae mortality was obtained with 25% A. cina and 75% T. lucida at 10 mg/mL. Additionally, this combination showed a synergistic effect. CONCLUSION: The two plant species studied here can be applied as natural anthelmintic alternatives due to their high bioactive effect and synergistic response.

Epidemiology and phylogeny of Haemonchus contortus through internal transcribed spacer 2 gene in small ruminants.

Introduction: Haemonchus contortus (H. contortus) is a blood-feeding nematode causing infectious disease haemonchosis in small ruminants of tropical and subtropical regions around the world. This study aimed to explore the prevalence and phylogeny of H. contortus in small ruminants using the internal transcribed spacer-2 (ITS-2) gene. In addition, a comprehensive review of the available literature on the status of H. contortus in Pakistan was conducted. Methods: Fecal samples were collected from sheep and goats (n = 180). Microscopically positive samples were subjected to DNA extraction followed by PCR using species-specific primers. Results: The overall prevalence of H. contortus was 25.55% in small ruminants. The prevalence of H. contortus was significantly associated with months and area. The highest occurrence of haemonchosis was documented in July (38.70%), whereas the lowest occurred in December (11.11%), with significant difference. The prevalence was highest in the Ghamkol camp (29.4%) and lowest in the arid zone of the Small Ruminant Research Institute (17.5%) (p = 0.01). The results of the systematic review revealed the highest prevalence of haemonchosis (34.4%) in Khyber Pakhtunkhwa (p = 0.001). Discussion: Phylogenetic analysis revealed a close relationship between H. contortus and isolates from Asia (China, India, Iran, Bangladesh, Malaysia, and Mongolia) and European countries (Italy and the United Kingdom). It has been concluded that H. contortus is prevalent in small ruminants of Kohat district and all over Pakistan, which could be a potential threat to food-producing animals, farmers, dairy, and the meat industry. Phylogenetic analysis indicates that H. contortus isolates share close phylogenetic relationships with species from Asia and Europe.

Comparative evolutionary analyses of peste des petits ruminants virus genetic lineages.

Peste des petits ruminants virus (PPRV) causes a highly infectious disease affecting mainly goats and sheep in large parts of Africa, Asia, and the Middle East and has an important impact on the global economy and food security. Full genome sequencing of PPRV strains has proved to be critical to increasing our understanding of PPR epidemiology and to inform the ongoing global efforts for its eradication. However, the number of full PPRV genomes published is still limited and with a heavy bias towards recent samples and genetic Lineage IV (LIV), which is only one of the four existing PPRV lineages. Here, we generated genome sequences for twenty-five recent (2010-6) and seven historical (1972-99) PPRV samples, focusing mainly on Lineage II (LII) in West Africa. This provided the first opportunity to compare the evolutionary pressures and history between the globally dominant PPRV genetic LIV and LII, which is endemic in West Africa. Phylogenomic analysis showed that the relationship between PPRV LII strains was complex and supported the extensive transboundary circulation of the virus within West Africa. In contrast, LIV sequences were clearly separated per region, with strains from West and Central Africa branched as a sister clade to all other LIV sequences, suggesting that this lineage also has an African origin. Estimates of the time to the most recent common ancestor place the divergence of modern LII and LIV strains in the 1960s-80s, suggesting that this period was particularly important for the diversification and spread of PPRV globally. Phylogenetic relationships among historical samples from LI, LII, and LIII and with more recent samples point towards a high genetic diversity for all these lineages in Africa until the 1970s-80s and possible bottleneck events shaping PPRV's evolution during this period. Molecular evolution analyses show that strains belonging to LII and LIV have evolved under different selection pressures. Differences in codon usage and adaptative selection pressures were observed in all viral genes between the two lineages. Our results confirm that comparative genomic analyses can provide new insights into PPRV's evolutionary history and molecular epidemiology. However, PPRV genome sequencing efforts must be ramped up to increase the resolution of such studies for their use in the development of efficient PPR control and surveillance strategies.

Determining Appropriate Numbers and Times of Daily Measurements Using GreenFeed System to Estimate Ruminal Methane Emission of Meat Goats.

The study was conducted to determine appropriate numbers and times of daily gas measurements to estimate total daily methane (CH4) emission of meat goats using a GreenFeed system (GFS). A replicated 4 (four measurement protocols) × 4 (four periods) Latin square design was employed with 16 Boer wethers in a confinement pen setting. Measurement protocols entailed three (G-3T; 0600-0700, 1400-1500, and 2200-2300 h), four (G-4T; 0700-0800, 1300-1400, 1900-2000, and 0100-0200 h), and six (G-6T; 0800-0900, 1200-1300, 1600-1700, 2000-2100, 0000-0100, and 0400-0500 h) times for daily measurement periods in GFS. The fourth protocol was continuous measurement over 24 h with animals in an open-circuit respiration calorimetry system (CS). Oat hay was given in individual feeders, and a small predetermined quantity of a pelleted concentrate supplement (bait) was dispensed by the GFS or manually offered for the CS. Overall, total dry matter (DM) intake (614, 625, 635, and 577 g/day for CS, G-3T, G-4T, and G-6T, respectively; SEM = 13.9) and digestible DM intake (359, 368, 374, and 320 CS, G-3T, G-4T, and G-6T, respectively; SEM = 15.9) were lower for CS than for G-3T, G-4T, and G-6T (p < 0.05), but these variables were not different among the GFS protocols. There was a significant (p < 0.001) effect of measurement protocol on CH4 emission in g/day (11.1, 25.6, 27.3, and 26.7 for CS, G-3T, G-4T, and G-6T, respectively; SEM = 1.11), g/kg DM intake (19.3, 46.4, 43.9, and 42.4 for CS, G-3T, G-4T, and G-6T, respectively; SEM = 2.03), and g/kg body weight (0.49, 1.11, 1.18, and 1.16 for CS, G-3T, G-4T, and G-6T, respectively; SEM = 0.052), with values being much lower for CS than for G-3T, G-4T and G-6T. Conversely, CH4 emission was similar among the GFS protocols despite differences in the time and number of daily visits (2.03, 2.76, and 3.75 visits for G-3T, G-4T, and G-6T, respectively; SEM = 0.114; p < 0.001). Pearson correlation (r) analysis indicated a moderate to high (p < 0.05) correlation between CS and G-3T (r = 0.62 for CH4 in g/day and r = 0.59 for CH4 in g/kg BW), CS and G-4T (r = 0.67 for CH4 in g/day and r = 0.76 for CH4 in g/kg BW), and CS and G-6T (r = 0.70 for CH4 in g/day and r = 0.75 for CH4 in g/kg BW). However, the correlation coefficient for CH4 in g/kg DM intake was low between CS and G-3T (r = 0.11) and CS and G-6T (r = 0.31) but slightly greater between CS and G-4T (r = 0.47). In conclusion, the results suggest that CH4 emissions using GFS in a confinement setting were greater compared with the CS in goats, but CH4-emission estimation using the GFS correlated with the CH4 emission in the CS system with a stronger relationship for the four times of daily measurements.

Animal Exposure Model for Mapping Crimean-Congo Hemorrhagic Fever Virus Emergence Risk.

To estimate the determinants of spatial variation in Crimean-Congo hemorrhagic fever virus (CCHFV) transmission and to create a risk map as a preventive public health tool, we designed a survey of small domestic ruminants in Andalusia, Spain. To assess CCHFV exposure spatial distribution, we analyzed serum from 2,440 sheep and goats by using a double-antigen ELISA and modeled exposure probability with environmental predictors by using generalized linear mixed models. CCHFV antibodies detected in 84 samples confirmed low CCHFV prevalence in small domestic ruminants in the region. The best-fitted statistical model indicated that the most significant predictors of virus exposure risk were cattle/horse density and the normalized difference vegetation index. Model validation showed 99.7% specificity and 10.2% sensitivity for identifying CCHFV circulation areas. To map CCHFV exposure risk, we projected the model at a 1 × 1-km spatial resolution. Our study provides insight into CCHFV ecology that is useful for preventing virus transmission.