Functionally distinct T-helper cell phenotypes predict resistance to different types of parasites in a wild mammal.

The adaptive immune system is critical to an effective response to infection in vertebrates, with T-helper (Th) cells pivotal in orchestrating these responses. In natural populations where co-infections are the norm, different Th responses are likely to play an important role in maintaining host health and fitness, a relationship which remains poorly understood in wild animals. In this study, we characterised variation in functionally distinct Th responses in a wild population of Soay sheep by enumerating cells expressing Th-subset specific transcription factors and quantifying Th-associated cytokines. We tested the prediction that raised Th1 and Th2 responses should predict reduced apicomplexan and helminth parasite burdens, respectively. All measures of Th-associated cytokine production increased with age, while Th17- and regulatory Th-associated cytokine production increased more rapidly with age in males than females. Independent of age, sex, and each other, IL-4 and Gata3 negatively predicted gastro-intestinal nematode faecal egg count, while IFN-γ negatively predicted coccidian faecal oocyst count. Our results provide important support from outside the laboratory that Th1 and Th2 responses predict resistance to different kinds of parasites, and illustrate how harnessing specific reagents and tools from laboratory immunology will illuminate our understanding of host-parasite interactions in the wild.

Genome-wide association study and genomic heritabilities for blood protein levels in Lori-Bakhtiari sheep.

Serum protein levels are related to physiological and pathological status of animals and could be affected by both genetic and environmental factors. This study aimed to evaluate genetic variation of serum protein profile in sheep. Blood samples were randomly collected from 96 Lori-Bakhtiari ewes, a heavy meat-type breed. Total protein, albumin, globulin, α1, α2, ß and γ globulins and IgG levels were measured in blood serum. The samples were genotyped using the Illumina OvineSNP50 BeadChip. The studied traits adjusted for age, birth type, birth season and estimate of breeding value for body weight were considered as pseudo-phenotypes in genome-wide association analysis. In the GWAS model, the first five principal components were fitted as covariates to correct the biases due to possible population stratification. The Plink, R and GCTA software were used for genome-wide association analysis, construction of Q-Q and Manhattan plots and estimation of genetic variances, respectively. Noticeable genomic heritabilities ± SE were estimated for total and γ globulins (0.868 ± 0.262 and 0.831 ± 0.364, respectively), but other protein fractions had zero or close to zero estimates. Based on the Bonferroni adjusted p values, four QTLs located on 181.7 Mbp of OAR3, 107.7 Mbp of OAR4, 86.3 Mbp of OAR7 and 83.0 Mbp of OAR8 were significantly associated with α1, ß, ß and γ globulins, respectively. The results showed that the PKP2, IGF2R, SLC22A1 and SLC22A2 genes could be considered as candidate genes for blood serum proteins. The present study showed significant genetic variations of some blood protein fractions.

Serological investigation and genotyping of Mycobacterium avium subsp. paratuberculosis in sheep and goats in Inner Mongolia, China.

Paratuberculosis a contagious and chronic disease in domestic and wild ruminants, is caused by Mycobacterium avium subspecies paratuberculosis (MAP). Typical clinical signs include intractable diarrhea, progressive emaciation, proliferative enteropathy, and mesenteric lymphadenitis. Paratuberculosis is endemic to many parts of the world and responsible for considerable economic losses. In this study, different types of paratuberculosis and MAP in sheep and goats were investigated in Inner Mongolia, a northern province in China contiguous with two countries and eight other provinces. A total of 4434 serum samples were collected from six cities in the western, central, and eastern regions of Inner Mongolia and analyzed using the ELISA test. In addition, tissue samples were collected from seven animals that were suspected to be infected with MAP. Finally, these tissues samples were analyzed by histopathological examination followed by polymerase chain reaction (PCR), IS1311 PCR-restriction enzyme analysis (PCR-REA), and a sequence analysis of five genes. Among all 4434 ruminant serum samples collected from the six cities in the western, central, and eastern regions of Inner Mongolia, 7.60% (337/4434) measured positive for the MAP antibody. The proportions of positive MAP antibody results for serum samples collected in the western, central, and eastern regions were 5.10% (105/2058), 6.63% (85/1282), and 13.44% (147/1094), respectively. For the seven suspected infected animals selected from the herd with the highest rate of positivity, the gross pathology and histopathology of the necropsied animals were found to be consistent with the pathological features of paratuberculosis. The PCR analysis further confirmed the diagnosis of paratuberculosis. The rest of the results demonstrated that herds of sheep and goats in Inner Mongolia were infected with both MAP type II and type III. To the best of our knowledge, this is the first study of the two subtypes of MAP strains in sheep and goats in Inner Mongolia.

Large-Scale International Validation of an Indirect ELISA Based on Recombinant Nucleocapsid Protein of Rift Valley Fever Virus for the Detection of IgG Antibody in Domestic Ruminants.

Diagnostic performance of an indirect enzyme-linked immunosorbent assay (I-ELISA) based on a recombinant nucleocapsid protein (rNP) of the Rift Valley fever virus (RVFV) was validated for the detection of the IgG antibody in sheep (n = 3367), goat (n = 2632), and cattle (n = 3819) sera. Validation data sets were dichotomized according to the results of a virus neutralization test in sera obtained from RVF-endemic (Burkina Faso, Democratic Republic of Congo, Mozambique, Senegal, Uganda, and Yemen) and RVF-free countries (France, Poland, and the USA). Cut-off values were defined using the two-graph receiver operating characteristic analysis. Estimates of the diagnostic specificity of the RVFV rNP I-ELISA in animals from RVF-endemic countries ranged from 98.6% (cattle) to 99.5% (sheep) while in those originating from RVF-free countries, they ranged from 97.7% (sheep) to 98.1% (goats). Estimates of the diagnostic sensitivity in ruminants from RVF-endemic countries ranged from 90.7% (cattle) to 100% (goats). The results of this large-scale international validation study demonstrate the high diagnostic accuracy of the RVFV rNP I-ELISA. Standard incubation and inactivation procedures evaluated did not have an adverse effect on the detectable levels of the anti-RVFV IgG in ruminant sera and thus, together with recombinant antigen-based I-ELISA, provide a simple, safe, and robust diagnostic platform that can be automated and carried out outside expensive bio-containment facilities. These advantages are particularly important for less-resourced countries where there is a need to accelerate and improve RVF surveillance and research on epidemiology as well as to advance disease control measures.

Antimicrobial Peptide Modifications against Clinically Isolated Antibiotic-Resistant Salmonella.

Antimicrobial peptides are promising molecules to address the global antibiotic resistance problem, however, optimization to achieve favorable potency and safety is required. Here, a peptide-template modification approach was employed to design physicochemical variants based on net charge, hydrophobicity, enantiomer, and terminal group. All variants of the scorpion venom peptide BmKn-2 with amphipathic α-helical cationic structure exhibited an increased antibacterial potency when evaluated against multidrug-resistant Salmonella isolates at a MIC range of 4-8 µM. They revealed antibiofilm activity in a dose-dependent manner. Sheep red blood cells were used to evaluate hemolytic and cell selectivity properties. Peptide Kn2-5R-NH2, dKn2-5R-NH2, and 2F-Kn2-5R-NH2 (variants with +6 charges carrying amidated C-terminus) showed stronger antibacterial activity than Kn2-5R (a variant with +5 charges bearing free-carboxyl group at C-terminus). Peptide dKn2-5R-NH2 (d-enantiomer) exhibited slightly weaker antibacterial activity with much less hemolytic activity (higher hemolytic concentration 50) than Kn2-5R-NH2 (l-enantiomer). Furthermore, peptide Kn2-5R with the least hydrophobicity had the lowest hemolytic activity and showed the highest specificity to Salmonella (the highest selectivity index). This study also explained the relationship of peptide physicochemical properties and bioactivities that would fulfill and accelerate progress in peptide antibiotic research and development.

Accumulation of PFAS by livestock - determination of transfer factors from water to serum for cattle and sheep in Australia.

In this study accumulation has been determined of several per- and polyfluoroalkyl substances (PFAS) from ingested water to steady state serum concentration for adult beef cattle and sheep raised on a hobby farm impacted by PFAS contamination. PFAS concentrations in stock water were stable for more than a year, they were non-measurable in grass but present at very low levels in soil which equated to just 1% of the intake from water. Prior to quantifying PFAS in cattle serum there had been no breeding for 18 months. Although there were high concentrations of several PFAS in the water, only perfluorooctane sulphonate (PFOS) and perfluorohexane sulphonate (PFHxS) were in cattle serum in appreciable amounts; perfluoroheptane sulphonate (PFHpS), perfluorononanoic acid (PFNA) and perfluorodecanoic acid (PFDA) were much lower. Transfer factors (TFs) for cattle were calculated by dividing steady state serum concentration (ng PFAS/mL) by water concentration (µg PFAS/L). Average and upper estimate TF values for cattle were calculated; the former were 140 (total PFOS, i.e. tPFOS), 130 (PFHpS), 65 (PFHxS), 170 (PFNA), and 120 (PFDA). Previous investigation campaigns at the farm provided relative steady state serum PFAS concentrations for sheep and cattle that allowed adjustment of the cattle TFs. The resulting average estimate TFs for ewes (non-pregnant and not lactating) were 20 (tPFOS) and 30 (PFHxS), other PFAS were not measurable in sheep serum. Discussion on using these TFs in human health risk assessments is provided. With certain assumptions/caveats the TFs allow estimations of PFAS steady state serum concentrations for use in preliminary human health risk assessments (HHRAs) when only PFAS in stock water is known.

Effects of fetal number on acute phase proteins, cortisol, and hematological parameters in ewes during the periparturient period.

Many subclinical diseases associated with inflammation occur in sheep during the periparturient period. Clinical symptoms are usually detected at an advanced stage of the disease; therefore, there are considerable risks of permanent health disorders in fetuses or dams. Determination of acute phase proteins (APPs) as markers of inflammation may allow for an earlier diagnosis and effective treatment. Furthermore, multi-fetus pregnancies are often associated with hematological disturbances. The study objective was to compare plasma concentrations of serum amyloid A (SAA), haptoglobin (Hp), fibrinogen (Fb), and cortisol in ewes bearing one and two fetuses in the period from 2 weeks before to 2 weeks after parturition as well as to determine hematological parameters in peripheral blood. There was an important effect of fetal number on APP and cortisol concentrations in periparturient ewes. There was a greater concentration of SAA, Hp, Fb, and cortisol in ewes bearing two fetuses compared with those bearing one fetus. Profiles for APP and cortisol concentrations and hematological parameters were similar for ewes bearing one and two fetuses, and trends were within normal reference ranges for the periparturient period. Furthermore, there were no differences in values for hematological variables between ewes bearing one and two fetuses. with there being no ewes with anemia. In summary, separate determination of the previously undefined physiological ranges of APPs and cortisol for ewes bearing one and two fetuses may facilitate diagnosis of subclinical disorders and enable comparison of laboratory test results with different reference values for ewes bearing different numbers of fetuses.

Physiological and hematological parameters of sheep reared in the tropics and subtropics / [Parâmetros fisiológicos e hematológicos de ovinos criados nos trópicos e nos subtrópicos]

This study aimed to determine the physiological and hematological parameters of sheep in Brazil. Therefore, 5,081 observations were used from previous experiments with animals from different Brazilian regions with the time of day, gender, breed, age, physiological characteristics, hematological and climatic variables. The animals were classified according to their stress level, and only data of non-stressed animals were used to calculate the reference values. Respiratory rate was the parameter that most differed from the normal range, with values ​​above the limits usually reported for the species, being higher in commercial wool sheep. For hematological variables, differences were observed between the present results and the previously reported values in the literature. A higher number of red blood cells (RBC), lower hemoglobin and mean corpuscular hemoglobin concentration (MCHC) were observed in young animals. A higher number of RBC and MCHC were noted in sheep crossed with local breeds. Considering that physiological and hematological parameters are indicators of the animal's pathological or physiological state, for a correct interpretation of the results, the use of reference values based on animals with similar racial characteristics and subjected to similar environmental conditions is recommended.(AU)

O objetivo deste estudo foi determinar parâmetros fisiológicos e hematológicos de ovinos no Brasil. Portanto, foram utilizadas 5.081 observações de experimentos anteriores com animais de diversas regiões do Brasil, com diferentes horário, sexo, raça, idade, características fisiológicas, variáveis ​​hematológicas e climáticas. Os animais foram classificados de acordo com o nível de estresse, e apenas os dados dos animais não estressados ​​foram utilizados para o cálculo dos valores de referência. A frequência respiratória foi o parâmetro que mais diferiu em relação à normalidade, com valores acima dos limites usualmente relatados para a espécie, sendo maior em ovinos de lã comercial. Para as variáveis ​​hematológicas, também foram verificadas diferenças entre os resultados observados e os valores previamente relatados na literatura. Maior número de hemácias, menor hemoglobina e concentração de hemoglobina corpuscular média (CHbCM) foram observados em animais jovens e maior número de hemácias e CHbCM em ovinos cruzados com raças locais. Considerando que os parâmetros fisiológicos e hematológicos são indicadores do estado patológico ou fisiológico do animal, para uma correta interpretação dos resultados recomenda-se a utilização de valores de referência baseados em animais com características raciais semelhantes e submetidos a condições ambientais também semelhantes.(AU)

Sheep Erythrocyte Preparation for Hemolytic Tests Exploring Complement Functional Activities.

Sheep erythrocytes (SE) are commonly used in complement functional tests. Non sensitized SE are useful to study the FH activity of cell protection. Indeed, as the cell surface of sheep erythrocytes is rich in sialic acids, Factor H (FH) is able to bind on it and therefore they represent a model of nonactivating surface. Because of their high capacity of complement regulation SE need to be modified to explore other functionality of the complement pathways, like the Complement hemolytic 50 (CH50) or the AP C3 convertase decay assays. For these tests, SE are sensitized with an anti-sheep red blood cell stroma antibody. In presence of serum or plasma complement components, sensitized SE may initiate complement cascade activation via the classic pathway explored in the CH50 assay. Sensitized SE may also be used to prepare C3b-coated SE that, with the use of buffers favoring AP, are suitable for the C3 Nef hemolytic assay and for the hemolytic assay studying the AP decay activity of FH. In this chapter we describe how to prepare SE for these different hemolytic tests.

Detection of C3 Nephritic Factor by Hemolytic Assay.

C3 nephritic Factor (C3NeF) is autoantibody that binds neoepitopes of the C3 convertase C3bBb, resulting in a stabilization of the enzyme. First functional characterizations of C3NeF were performed by hemolytic assays using preactivated sheep erythrocytes (bearing C3b). Sheep erythrocytes are beforehand sensitized with an anti-sheep red blood cell stroma antibody produced in rabbit (hemolysin). Sensitized sheep erythrocytes will initiate cascade complement activation via the classic pathway, followed by alternative pathway amplification loop, resulting in C3b covalent binding to cell surface. Sheep erythrocytes bearing C3b permit the alternative pathway exploration, in particular decay of alternative pathway C3 convertase.

Role of eprinomectin as inhibitor of the ruminant ABCG2 transporter: Effects on plasma distribution of danofloxacin and meloxicam in sheep.

Therapeutic outcome results of the coadministration of several drugs in veterinary medicine is affected by, among others, the relationship between drugs and ATP-binding cassette (ABC) transporters, such as ABCG2. ABCG2 is an efflux protein involved in the bioavailability and milk secretion of drugs. The aim of this work was to determine the role of eprinomectin, a macrocyclic lactone (ML) member of avermectin class, as inhibitor of ABCG2. The experiments were carried out through in vitro inhibition assays based on mitoxantrone accumulation and transport assays in ovine ABCG2 transduced cells using the antimicrobial drug danofloxacin and the anti-inflammatory drug meloxicam, both widely used in veterinary medicine and well known ABCG2 substrates. The inhibition results obtained showed that eprinomectin was an efficient in vitro ABCG2 inhibitor, tested in mitoxantrone accumulation assays. In addition, this ML decreased ovine ABCG2-mediated transport of danofloxacin and meloxicam. To evaluate the role of eprinomectin in systemic exposure of drugs, pharmacokinetic assays based on subcutaneous coadministration of eprinomectin with danofloxacin (1.25 mg/kg) or meloxicam (0.5 mg/kg) in sheep were performed obtaining a significant increase of systemic exposure of these drugs. Especially relevant was the increase of the systemic concentration of meloxicam, since coadministration with eprinomectin increased significantly the plasma concentration of meloxicam, obtaining an increase of AUC (0-72 h) value of more than 40%.

A centrifugal microfluidic cross-flow filtration platform to separate serum from whole blood for the detection of amphiphilic biomarkers.

The separation of biomarkers from blood is straightforward in most molecular biology laboratories. However, separation in resource-limited settings, allowing for the successful removal of biomarkers for diagnostic applications, is not always possible. The situation is further complicated by the need to separate hydrophobic signatures such as lipids from blood. Herein, we present a microfluidic device capable of centrifugal separation of serum from blood at the point of need with a system that is compatible with biomarkers that are both hydrophilic and hydrophobic. The cross-flow filtration device separates serum from blood as efficiently as traditional methods and retains amphiphilic biomarkers in serum for detection.

Effect of climatic factors on urinary cortisol and peripheral blood leukocytes in lambs grazing on a semi-natural grassland in the Hokuriku District of Japan.

The effects of climatic factors on stress and immune functions of grazing lambs in summer and autumn in the Hokuriku District of Japan were evaluated by determining urinary cortisol (U-COR) levels and peripheral blood leukocyte populations and comparing those with lambs kept indoors. Two groups of five lambs, consisting of those grazed on a semi-natural grassland (GRL) and those housed indoors in a domestic animal shelter (INL), were maintained at from July to October. The temperature-humidity index at each location was indicative of heat stress during summer; however, the U-COR elevation was not observed in both groups. The elevation was observed in GRL in autumn and was higher than INL in October. Climatic conditions in autumn were characterized by high humidity and a sudden drop in temperature. U-COR was positively correlated with the relative humidity. The GRL was exposed to low-nutrient conditions for a relatively long time. The CD4+ /CD8+ T cell ratio in GRL decreased in October. Subsequently, the total leucocyte, including granulocytes, monocytes, and lymphocytes, sharply increased. The responses indicated an immune deficiency caused by immunosuppression because of a low nutrition caused by grazing and high-stress conditions in autumn.

Differences in mean grey levels of uterine ultrasonographic images between non-pregnant and pregnant ewes may serve as a tool for early pregnancy diagnosis.

The aim of this study was to determine how physiological and hormonal changes in the uterus during the estrous cycle and early gestational period affect the average grey values of pixels when conducting computer-assisted analysis of uterine ultrasonic images in ewes. For this purpose, 60 ewes on which there had been an estrous synchrony regimen imposed were included in the study. Animals were assigned to two groups with ewes not being mated and assessments occurred during the subsequent estrous cycle (Group 1; n = 25) and ewes being mated and assessments occurring during the subsequent early gestational period (Group 2; n = 35). Ewes were examined utilizing real time ultrasonic procedures and uterine images were obtained. Digital analysis of uterine ultrasonographic images was performed using image J program and mean grey levels (MGL) were determined. Values for progesterone concentrations were consistent with those previously reported in non-pregnant and pregnant ewes. There was a close association between MGL values in ewes of both Group I (P < 0.05) and II (P < 0.05) and days of the estrous cycle. There was also an association between MGL values and day of the gestational period in ewes of Group 2(P < 0.001). In conclusion, there are differences in MGL values between non-pregnant and pregnant ewes with there being changes as days of the estrous cycle and day of gestation period advances, therefore, this procedure could be used as a pregnancy diagnostic criterion during the early period of gestation in ewes.

Hematologic Conditions of Small Ruminants.

Anemia is a clinically important syndrome in small ruminants. Anemia can be divided into regenerative and nonregenerative forms. Differentials for regenerative anemia include hemorrhage owing to gastrointestinal or external parasitism or hemostatic disorders, and hemolysis owing to infectious, osmotic, toxic, and nutritional causes. Differentials for nonregenerative anemia include inflammatory and chronic diseases, renal failure, pancytopenia, copper deficiency, and heavy metal toxicosis. Iron deficiency anemia can be caused by chronic gastrointestinal and external hemorrhage or nutritional deficiency and may be mildly regenerative or nonregenerative. Appropriate diagnostic tests are described along with treatments, including blood transfusion, parasite control, and prevention.

Effect of date palm (Phoenix dactylifera L.) leaves on productive performance of growing lambs.

Eighteen 4-month-old lambs, with a mean live weight (LW) of 19.47 ± 0.20 kg, were used to evaluate the nutritive value of date palm leaves (DPL) ensiled with different additives in a completely randomized design. Lambs were stratified into three groups of 6 lambs each and fed a control diet comprising 60% concentrate feed mixture (CFM) and 40% DPL silage (T1). In other treatments, the DPL silage (DPLS) of the control treatment was replaced with EM1 additive-treated DPLS (T2) or El-Mofeed additive-treated DPLS (T3). Apparent digestibility, total digestible nutrient, digestible crude protein, dry matter intake, daily weight gain (DWG), price of DWG, daily profit, and economics of feed efficiency were higher (P < 0.05) for the additives-treated DPLS relative to the control, with T2 enhancing these parameters compared with T3. With exception of ruminal pH, which was reduced, concentrations of ruminal NH3-N and total volatile fatty acids (VFA) increased 4 h post feeding. However, ruminal NH3-N and total VFA were greater (P < 0.05) for the additives-treated DPLS, with T2 producing higher values than T3. Ruminal pH and feed cost/kg LW gain were lower for T2 relative to other treatments. Blood constituents were within the normal ranges for lambs, though slightly altered by treatments. Whereas serum total protein, albumin, and globulin were affected (P < 0.05) in this rank order, T1 < T3 < T2, other serum parameters were not affected. Relative feed cost and relative daily profit were lower and higher respectively for T2 than for T3. It is concluded that additives-treated DPLS is nutritionally superior to untreated DPLS as a roughage source in total mixed rations fed to growing lambs. However, for improved performance of the lambs and economic benefits, EM1-treated DPLS is recommended.

Novel circRNA discovery in sheep shows evidence of high backsplice junction conservation.

Circular RNAs (circRNAs) are covalently closed circular non-coding RNAs. Due to their structure, circRNAs are more stable and have longer half-lives than linear RNAs making them good candidates for disease biomarkers. Despite the scientific relevance of these molecules, the study of circRNAs in non-model organisms is still in its infancy. Here, we analyse total RNA-seq data to identify circRNAs in sheep from peripheral blood mononuclear cells (PBMCs) and parietal lobe cortex. Out of 2510 and 3403 circRNAs detected in parietal lobe cortex and in PBMCs, a total of 1379 novel circRNAs were discovered. Remarkably, around 63% of all detected circRNAs were found to be completely homologous to a circRNA annotated in human. Functional enrichment analysis was conducted for both tissues based on GO terms and KEGG pathways. The enriched terms suggest an important role of circRNAs from encephalon in synaptic functions and the involvement of circRNAs from PBMCs in basic immune system functions. In addition to this, we investigated the role of circRNAs in repetitive vaccination experiments via differential expression analysis and did not detect any significant relationship. At last, our results support both the miRNA sponge and the miRNA shuttle functions of CDR1-AS in sheep brain. To our knowledge, this is the first study on circRNA annotation in sheep PBMCs or parietal lobe cortex samples.

The common European mosquitoes Culex pipiens and Aedes albopictus are unable to transmit SARS-CoV-2 after a natural-mimicking challenge with infected blood.

BACKGROUND: On 11 March 2020, the World Health Organisation (WHO) declared the coronavirus disease 2019 (COVID-19) outbreak to be a pandemic. As the mosquito season progressed, the understandable concern that mosquitoes could transmit the virus began to increase among the general public and public health organisations. We have investigated the vector competence of Culex pipiens and Aedes albopictus, the two most common species of vector mosquitoes in Europe, for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Due to the very unusual feeding behaviour of Ae. albopictus, we also evaluated the role of this mosquito in a potential mechanical transmission of the virus. METHODS: For the vector competence study, mosquitoes were allowed to take several infectious blood meals. The mosquitoes were then collected and analysed at 0, 3, 7 and 10 days post-feeding. For the mechanical transmission test, Ae. albopictus females were allowed to feed for a short time on a feeder containing infectious blood and then on a feeder containing virus-free blood. Both mosquitoes and blood were tested for viral presence. RESULTS: Culex pipiens and Ae. albopictus were found not be competent vectors for SARS-CoV-2, and Ae. albopictus was unable to mechanically transmit the virus. CONCLUSIONS: This is the first study to show that the most common species of vector mosquitoes in Europe do not transmit SARS-CoV-2 and that Ae. albopictus is unable to mechanically transmit the virus from a positive host to a healthy host through host-feeding.

Pharmacokinetic comparison of six anthelmintics in sheep, goats, and cattle.

This study was initiated to determine whether a comparative pharmacokinetic (PK) approach could be used to expand the pool of approved anthelmintics for minor ruminant species. Accordingly, the PK profiles of six anthelmintics (levamisole, albendazole, fenbendazole, moxidectin, doramectin, and ivermectin) in sheep, goats, and cattle were determined. The PK values determined for each anthelmintic included Tmax , Tlast , Cmax , AUC, AUC/dose, and Cmax /dose. The results of this study demonstrate that a comparative PK approach does not show commonality in the way these six anthelmintics are individually processed by these three ruminants. While some drugs demonstrated identical PK profiles between sheep and goats, none of these drugs demonstrated PK profiles in sheep and goats comparable to the PK profiles found in cattle. The results from this study suggest drug approval across these three ruminants is not a viable concept. However, the resulting PK profiles for each combination of drug and ruminant species represents a new dataset that can be used to support the US FDA Center for Veterinary Medicine's Minor Use/Minor Species indexing process for drug approvals in minor species such as sheep and goats.

Ovine red cell concentrates for transfusion research - is the storage lesion comparable to human red cell concentrates?

BACKGROUND AND OBJECTIVES: Sheep are increasingly being used as a large in vivo animal model of blood transfusion because they provide several advantages over small animals. Understanding the effects of storage duration on ovine (ov) red cell concentrates (RCCs) and how these changes compare with stored human (hu) RCCs is necessary to facilitate clinical translation of research findings. MATERIALS AND METHODS: OvRCCs (n = 5) collected and processed in standard human blood collection packs, and equivalent huRCCs provided by Australian Red Cross Lifeblood (n = 5), were stored at 2-6°C for 42 days, with samples collected weekly. Haemolysis index was determined by measuring supernatant haemoglobin concentration. Biochemical parameters were evaluated using a blood gas analyser. Energy metabolites and biologically active lipids were measured using commercial assays. Osmotic fragility was determined by lysis in various saline concentrations. Extracellular vesicles were characterized by nanoparticle tracking analysis. RESULTS: Ovine red blood cells (RBCs) are double in number, smaller in size and more fragile than human RBCs. Haematological values were unchanged throughout storage. In contrast, biochemical and metabolic values, and haemolysis index in three of the five ovRCCs exceeded huRCCs licensing criteria by day 42. Accumulation of extracellular vesicles and biologically active lipids was comparable between huRCCs and ovRCCs. CONCLUSION: This study documents similarities and differences in the storage lesion of ovRCCs and huRCCs. This new information will guide the design of ovine transfusion models to enhance translation of findings to human transfusion settings.