Cellular composition of granulomatous lesions in gut-associated lymphoid tissues of goats during the first year after experimental infection with Mycobacterium avium subsp. paratuberculosis

Mycobacterium avium subsp. paratuberculosis (MAP) causes lesions in naturally and experimentally infected ruminants which greatly differ in severity, cellular composition and number of mycobacteria. Morphologically distinct lesions are already found during the clinically inapparent phase of infection. The complex local host response and number of MAP were characterized at the initial sites of lesions, organized gut-associated lymphoid tissue, in experimentally infected goats. Tissues were collected at 3, 6, 9 and 12 month post-inoculation (mpi) from goat kids that had orally received 10 times 10 mg of bacterial wet mass of MAP (JII-1961). The cellular composition of lesions in Peyer's patches in the jejunum and next to the ileocecal valve was evaluated in 21 MAP-inoculated goats, where lesions were compared with unaltered tissue of six control goats. CD68⁺, CD4⁺, CD8⁺, γδ T lymphocytes, B lymphocytes and plasma cells, MHC class II⁺ and CD25⁺ cells were demonstrated by immunohistochemistry in serial cryostat sections.At 3 mpi, extensive granulomatous infiltrates predominated, consisting of numerous epitheloid cells admixed with many CD4 and γδ T lymphocytes. Only single MAP were detected. This indicates a strong cellular immune reaction able to control MAP infection. γδ T lymphocytes were markedly increased in this type of lesion which may reflect their important role early in the pathogenesis of paratuberculosis. At 9 and 12 mpi, divergent lesions were observed which may reflect different outcomes of host–pathogen interactions. In five goats, minimal granulomatous lesions were surrounded by extensive lymphoplasmacytic infiltrates and no MAP were detected by immunohistochemistry. This was interpreted as effective host response that was able to eliminate MAP locally. In three goats, decreased numbers of lymphocytes, but extensive granulomatous infiltrates with numerous epitheloid cells containing increased numbers of mycobacteria were seen. This shift of the immune response resulted in uncontrolled mycobacterial multiplication. Focal and multifocal circumscribed granulomatous infiltrates of mainly epitheloid cells may represent sites of new infection, since they were observed in goats at all times after inoculation. Their presence in goats with minimal granulomatous lesions surrounded by extensive lymphoplasmacytic infiltrates may indicate that despite the local clearance, the infection may be perpetuated.The complex cellular immune reactions postulated for the pathogenesis of paratuberculosis were demonstrated at the local sites of infection. These early host–pathogen interactions are most likely essential for the eventual outcome of the MAP infection.     

Evaluation of peripheral lymphocytes after weaning and vaccination for Mycoplasma hyopneumoniae

This study evaluated immune cell populations in pigs following weaning and vaccination for Mycoplasma hyopneumoniae. Piglets (n = 24) were weaned (day 0) at 16 (±1) days of age, and randomly assigned to the vaccination group (n = 16) or control group (n = 8). Complete blood cell counts, flow cytometry and serology were completed for blood samples collected on days 0 (within hours of weaning), 3, 7, 14, 30 and 60. The M. hyopneumoniae S:P ratios (sample optical density: positive control optical density) were negative in the vaccination group until days 30 and 60, when the S:P ratios were 1.3 and 1.0, respectively. Control animals remained serologically negative. The percentage of CD4⁺ T cells was less (P < 0.01) in control pigs than vaccinated pigs at day 3. In contrast, numbers of CD8⁺ and CD4⁺CD8⁺ T cells were greater (P < 0.01) in control pigs than in vaccinated pigs at days 3 and 7. After day 7, few differences in immune cell types were evident between the groups. Differences in lymphocyte populations could not be solely attributed to vaccination, due at least in part, to the confounding influence of weaning. It was difficult to distinguish the influence of vaccination from the impact of weaning on peripheral immune cell populations.     

Effect of supplemental sericea lespedeza leaf meal pellets on gastrointestinal nematode infection in grazing goats

Feeding sun-dried sericea lespedeza [SL; Lespedeza cuneata (Dum-Cours.) G. Don.] reduces gastrointestinal nematode (GIN) infection in goats fed in confinement, but effects of this forage when fed as a supplement to goats on pasture are unclear. A study was completed in which supplemental feeds (75 and 95% SL leaf meal pellets and a commercial pellet, all fed at 0.91 kg/head/day) were offered to thirty growing male Spanish goats (9 months old, 20.6 ± 2.8 kg, 10/treatment) grazing perennial warm-season grass pastures in Fort Valley, GA, from September to November, 2010. Fecal and blood samples were taken from individual animals weekly to determine fecal egg count (FEC) and packed cell volume (PCV), respectively, and animal weights were recorded at the start and end of the trial. After 11 weeks grazing, animals were slaughtered for recovery, counting, and speciation of adult GIN from the abomasum and small intestines. There was no difference in FEC between goats fed the 75 and 95% SL leaf meal pellets, but both groups had lower (P < 0.05) FEC than the goats fed the commercial pellets from days 35 to 77. The PCV values were not affected by the dietary treatments. Animal gain per day averaged 102.0, 77.2, and 53.3 g for goats fed 95% SL, commercial, and 75% SL pellets, respectively (P < 0.05). The 95% SL leaf meal pellet goats had 93.0 and 47.3% fewer (P < 0.05) total (male + female) adult Haemonchus contortus and Teladorsagia circumcincta, respectively, than control animals, while only male H. contortus were lower (47.6%; P < 0.05) in 75% SL-fed goats compared with commercial pellet-fed animals. Feeding supplemental SL leaf meal pellets improved animal performance (95% SL pellets) and reduced worm burdens (75 and 95% SL pellets) in young grazing goats and is a useful tool for natural GIN control in small ruminants.     

Replication of neurovirulent equine herpesvirus type 1 (EHV-1) in CD172a+ monocytic cells

Equine herpesvirus type 1 (EHV-1) is responsible for respiratory disorders, abortion and myeloencephalopathy (EHM) in horses. Two pathotypes of EHV-1 strains are circulating in the field: neurovirulent (N) and non-neurovirulent (NN). For both strains, CD172a⁺ monocytic cells are one of the main carrier cells of EHV-1 during primary infection, allowing the virus to invade the horse’s body. Recently, we showed that EHV-1 NN strains showed a restricted and delayed replication in CD172a⁺ cells. Here we characterize the in vitro replication kinetics of two EHV-1 N strains in CD172a⁺ cells and investigate if the replication of these strains is similarly silenced as shown for EHV-1 NN strains. We found that EHV-1 N replication was restricted to 7–8% in CD172a⁺ cells compared to 100% in control RK-13 cells. EHV-1 N replication was not delayed in CD172a⁺ cells but virus production was significant lower (10^3.0 TCID₅₀/10⁵ inoculated cells) than in RK-13 cells (10^8.5 TCID₅₀/10⁵ inoculated cells). Approximately 0.04% of CD172a⁺ cells produced and transmitted infectious EHV-1 to neighbour cells compared to 65% of RK-13 cells. Unlike what we observed for the NN strain, pretreatment of CD172a⁺ cells with histone deacetylases inhibitors (HDACi) did not influence the replication of EHV-1 N strains in these cells. Overall, these results show that the EHV-1 replication of N strains in CD172a⁺ cells differs from that observed for NN strains, which may contribute to their different pathogeneses in vivo.     

Phenotypic and functional properties of feline dedifferentiated fat cells and adipose-derived stem cells

It has been reported that mature adipocyte-derived dedifferentiated fat (DFAT) cells show multilineage differentiation potential similar to that observed in mesenchymal stem cells. Since DFAT cells can be prepared from a small quantity of adipose tissue, they could facilitate cell-based therapies in small companion animals such as cats. The present study examined whether multipotent DFAT cells can be generated from feline adipose tissue, and the properties of DFAT cells were compared with those of adipose-derived stem cells (ASCs). DFAT cells and ASCs were prepared from the floating mature adipocyte fraction and the stromal vascular fraction, respectively, of collagenase-digested feline omental adipose tissue. Both cell types were evaluated for growth kinetics, colony-forming unit fibroblast (CFU-F) frequency, immunophenotypic properties, and multilineage differentiation potential.DFAT cells and ASCs could be generated from approximately 1 g of adipose tissue and were grown and subcultured on laminin-coated dishes. The frequency of CFU-Fs in DFAT cells (35.8%) was significantly higher than that in ASCs (20.8%) at passage 1 (P1). DFAT cells and ASCs displayed similar immunophenotypes (CD44⁺, CD90⁺, CD105⁺, CD14^?, CD34^? and CD45^?). Alpha-smooth muscle actin-positive cells were readily detected in ASCs (15.2 ± 7.2%) but were rare in DFAT cells (2.2 ± 3.2%) at P1. Both cell types exhibited adipogenic, osteogenic, chondrogenic, and smooth muscle cell differentiation potential in vitro. In conclusion, feline DFAT cells exhibited similar properties to ASCs but displayed higher CFU-F frequency and greater homogeneity. DFAT cells, like ASCs, may be an attractive source for cell-based therapies in cats.     

Rough Agave Flowers as a Potential Feed Resource for Growing Goats

The objective of this study was to evaluate the effect of different levels of rough agave (Agave scabra Ortega) flowers on dry matter intake (DMI), average daily gain (ADG), volatile fatty acid (VFA) production in the rumen, and particular serum metabolites and minerals of native × dairy growing goats (Capra hircus L.). Forty female goats with an initial weight of 11.1 ±  kg (mean ± SD) were used in a completely randomized design experiment that lasted for 84 d. Goats were fed a completely mixed ration (30% roughage, 70% ground corn [Zea mays L.] and soybean [Glycine max {L.} Merr] meal). Treatments consisted of offering goats (4 pens · group⁻¹, 2 goats · pen⁻¹) air-dry rough agave flowers, which replaced alfalfa (Medicago sativa L.) hay at 0% (control; T0), 25% (T25), 50% (T50), 75% (T75), and 100% (T100) of the of the roughage portion of the diet. Values of nutritional parameters for rough agave flowers were in vitro organic matter digestibility, 493 g · kg⁻¹; crude protein, 115 g · kg⁻¹; and metabolizable energy, 6.29 MJ · kg⁻¹ DMI. There were differences (P < 0.05) in ADG (range, 108–155 g · d⁻¹) between diets. Goats fed T0 had higher (P < 0.05) gains than goats fed T50 and T100. DMI was not affected by dietary treatments (range, 3.4% to 3.6% of body weight). Feed conversion ratio (FCR, defined as DMI/ADG) increased (P < 0.05) 27% with total substitution of alfalfa by rough agave flowers, in comparison with T0. Lower (P < 0.05) values of total VFA were obtained with T100, in comparison with all other dietary treatments. These results demonstrated that totally replacing alfalfa with rough agave flowers in diets did not affect DMI but decreased AGD and compromised FCR. Thus, rough agave flowers have the potential to partially replace alfalfa in diets for growing goats.     

Improvements of immune status, intestinal integrity and gain performance in the early-weaned calves parenterally supplemented with L-alanyl-L-glutamine dipeptide

Glutamine (Gln) is an important substrate for the innate immune cells including lymphocytes and macrophages. In this report, the effects of alanyl-glutamine dipeptide (Ala-Gln) on the naïve immune system, intestinal integrity and gain performance of early-weaned calves were investigated. Early-weaned Chinese Holstein calves were intravenously administered different dosages of Ala-Gln. The effects of Ala-Gln administration were ascertained by evaluating the blood for naïve T lymphocyte subpopulations, the concentrations of serum IgG, serum IgA and intestinal mucosal secretory IgA (s-IgA), the intestinal integrity, as well as the gain performance. Results demonstrated that intravenous administration of Ala-Gln dipeptide (1.01 g/kg × d^?1) for 7 days had a positive effect on gain performance, intestinal integrity and the immune system. Calves administered doses of Ala-Gln displayed an improvement in gain performance and health status concurrent with increases in blood CD2⁺ and CD4⁺ lymphocytes, the ratio of CD4⁺/CD8⁺, serum IgA and IgG, intestinal mucosal s-IgA while decreasing the occurrence of diarrhea. Moreover, we found that animals given the effective dose (1.01 g/kg × d^?1) of Ala-Gln resulted in improved immune status and intestinal integrity relative to those given a lower (0.49 g/kg × d^?1) or higher dose (1.99 g/kg × d^?1) of Ala-Gln. These findings suggest that maintaining a certain concentration of plasma and/or tissue glutamine in the early stages of weaning is an effective alternative approach for improvement of growth performance in early-wean calves.     

Age and Body Condition of Goats Influence Consumption of Juniper and Monoterpene-Treated Feed

Redberry juniper (Juniperus pinchotii Sudworth) is an invasive, evergreen tree that is rapidly expanding throughout western and central Texas. Goats will consume some juniper on rangelands; however, intake is limited. The objective of our research was to determine how the age and body condition of goats influence their consumption of juniper and an artificial feed containing 4 monoterpenes. Two separate experiments were conducted. Experiment 1 examined the intake of redberry juniper foliage and used 39 goats either young (2 yr) or mature (> 6 yr). One-half of each age group was fed appropriate basal rations to reach either a high (HBC) or low body condition (LBC). Goats in LBC ate more (P < 0.01, 8.6 g · kg⁻¹ body weight [BW] ± 0.7 SE) juniper than those in HBC (2.3 g · kg⁻¹ BW ± 0.3 SE), and young animals consumed more (P < 0.05, 7.2 g · kg⁻¹ BW ± 0.7 SE) juniper than mature goats (3.9 g · kg⁻¹ BW ± 0.5 SE) across body condition treatments. In experiment 2, 36 goats, either young (2 yr) or mature (> 6 yr) and in either HBC or LBC, were offered a synthetic ration treated with 20.8 g · kg⁻¹ of 4 monoterpenes found in redberry juniper. Goats in LBC ate more (P < 0.01, 25.3 g · kg⁻¹ BW ± 1.0 SE) of the terpene-treated feed than those in HBC (17.5 g · kg⁻¹ BW ± 0.7 SE), and young animals ate more (P < 0.05, 22.5 g · kg⁻¹ BW ± 0.8 SE) than mature goats (20.3 g · kg⁻¹ BW ± 0.8 SE) across body condition treatments. Total intake as a proportion of body weight was also affected by body condition. Age and body condition are important factors that influence intake of chemically defended plants. A better understanding of how these attributes affect diet selection will aid livestock producers in improving grazing management.     

In vitro studies on the influence of dexamethasone and meloxicam on bovine WC1+ γδ T cells

In view of the lack of data on the effect of meloxicam (non-steroidal anti-inflammatory drug) on bovine γδ T cells (WC1⁺ cells) and very poorly recognized effects of dexamethasone (steroidal anti-inflammatory drug) on these cells, the purpose of the present study has been to determine the in vitro influence of these drugs on CD25^highWC1⁺, CD25^lowWC1⁺ and CD25^?WC1⁺ lymphocytes of the peripheral blood of cattle. Peripheral blood mononuclear cells were treated with the drugs in concentrations reflecting their plasma levels achieved in vivo at therapeutic doses (dexamethasone 10^?7 M; meloxicam 5 × 10^?6 M) and at ten-fold lower concentrations. It was found out that percentages and absolute counts of CD25^highWC1⁺ and CD25^lowWC1⁺ cells increased in the presence of dexamethasone, and this effect was at least partly attributable to lower mortality of these cells, whose apoptosis was depressed by exposure to dexamethasone. It seems certain that this effect was not a result of increased multiplication of CD25^highWC1⁺ and CD25^lowWC1⁺ cells because their proliferation was reduced in the presence of dexamethasone. Exposure to this drug caused a rapidly occurring and lasting depletion of CD25^?WC1⁺, which was at least partly due to their higher apoptosis. The results seem to suggest that impaired proliferation of these cells was responsible for a more profound expression of this disorder. Paradoxically, the percentage of cells producing IFN-γ, a proinflammatory cytokine, increased in the presence of dexamethasone, whereas the count of cells secreting the key anti-inflammatory and immunosuppressive cytokine, i.e. IL-10, declined. This effect was observed in all analyzed subpopulations of cells. Meloxicam did not interfere so drastically as dexamethasone with the functioning of WC1⁺ lymphocytes because it did not affect their apoptosis, proliferation, percentage or absolute count. With respect to the effect of meloxicam on counts of particular WC1⁺ lymphocyte subpopulations, it was only demonstrated that exposure to the drug was correlated with a transient and very weakly expressed decrease in the relative and absolute counts of CD25^highWC1⁺ and CD25^lowWC1⁺ cells, which was most probably a result of a temporary down-regulation of the expression of the CD25 molecule. In the presence of meloxicam, percentages of IFN-γ⁺CD25^?WC1⁺ cells as well as cells producing IL-10 declined, an effect observed in all analyzed cell populations. These results suggest that care should be taken when administering this medication to animals with bacterial or viral infections, and we should avoid giving it to patients suffering from allergic or autoimmune disorders.     

Mycoplasma ovis infection in goat farms from northeastern Brazil

Although Mycoplasma ovis (formerly Eperythrozoon ovis) has been described in small ruminants worldwide, data on M. ovis in goats remain scarce. Accordingly, the aims of the present study were to i) determine the prevalence of hemoplasmas in goats, ii) identify the tick species parasitizing the animals, and iii) determine factors associated with infection in five dairy and three beef goat farms from the Paraíba State, northeastern Brazil. Blood samples were obtained from 402 goats. Samples were screened for hemoplasmas using a pan-hemoplasma PCR. The positive samples were confirmed by sequencing. An epidemiological questionnaire was given to each farm owner addressing age, gender, and presence of ticks. A total of 158/402 (39.3%) goats were positive for M. ovis by PCR. Sequencing of PCR positive samples has shown ≥99% identity with multiple M. ovis 16S rDNA sequences deposited in GenBank, including M. ovis isolates from humans. Dairy (OR = 2.15; 95% CI: 1.40–3.32%; P = 0.0004) and anemic goats (OR = 2.33; 95% CI: 1.51–3.71%; P = 0.0001) were more likely to be infected than beef and non-anemic animals, respectively. Amblyomma parvum (49/52, 94.23%) and Rhipicephalus microplus (3/52, 5.77%) were the tick species found parasitizing the animals, with no significant association between the presence of ticks and infection by M. ovis (P = 0.1164). This is the first reportedly molecular detection of M. ovis infection in goats from South America. In conclusion, M. ovis is highly prevalent in goats from northeastern Brazil, mainly in dairy animals.     

Antibody and T cell responses induced in chickens immunized with avian influenza virus N1 and NP DNA vaccine with chicken IL-15 and IL-18

We had examined the immunogenicity of a series of plasmid DNAs which include neuraminidase (NA) and nucleoprotein (NP) genes from avian influenza virus (AIV). The interleukin-15 (IL-15) and interleukin-18 (IL-18) as genetic adjuvants were used for immunization in combination with the N1 and NP AIV genes. In the first trial, 8 groups of chickens were established with 10 specific-pathogen-free (SPF) chickens per group while, in the second trial 7 SPF chickens per group were used. The overall N1 enzyme-linked immunosorbent assay (ELISA) titer in chickens immunized with the pDis/N1 + pDis/IL-15 was higher compared to the chickens immunized with the pDis/N1 and this suggesting that chicken IL-15 could play a role in enhancing the humoral immune response. Besides that, the chickens that were immunized at 14-day-old (Trial 2) showed a higher N1 antibody titer compared to the chickens that were immunized at 1-day-old (Trial 1). Despite the delayed in NP antibody responses, the chickens co-administrated with IL-15 were able to induce earlier and higher antibody response compared to the pDis/NP and pDis/NP + pDis/IL-18 inoculated groups. The pDis/N1 + pDis/IL-15 inoculated chickens also induced higher CD8+ T cells increase than the pDis/N1 group in both trials (P < 0.05). The flow cytometry results from both trials demonstrated that the pDis/N1 + pDis/IL-18 groups were able to induce CD4+ T cells higher than the pDis/N1 group (P < 0.05). Meanwhile, pDis/N1 + pDis/IL-18 group was able to induce CD8+ T cells higher than the pDis/N1 group (P < 0.05) in Trial 2 only. In the present study, pDis/NP was not significant (P > 0.05) in inducing CD4+ and CD8+ T cells when co-administered with the pDis/IL-18 in both trials in comparison to the pDis/NP. Our data suggest that the pDis/N1 + pDis/IL-15 combination has the potential to be used as a DNA vaccine against AIV in chickens.     

Upfront denileukin diftitox as in vivo regulatory T-cell depletion in order to enhance vaccination effects in a canine allogeneic hematopoietic stem cell transplantation model

Denileukin Diftitox (ONTAK^®, DAB₃₈₉ IL-2) is a recombinant DNA-derived fusion protein depleting cells that express high-affinity IL-2 receptor. Important cell targets are CD4⁺CD25⁺Foxp3⁺ regulatory T cells (T_reg). Elimination of immunosuppressive T_reg by Denileukin Diftitox may provide a way to modulate immune tolerance following stem cell transplantation. Here, we combined T_reg depletion with a vaccination approach to induce donor-specific immune reactions. To investigate this approach we chose the mixed chimerism canine stem cell transplantation model which represents a high state of tolerance between two hematopoietic systems. The aim was therefore to induce a graft versus hematopoiesis effect thereby converting mixed to full donor chimerism. Dog leukocyte antigen identical siblings that had developed a stable mixed chimerism after non-myeloablative stem cell transplantation received a single dose of Denileukin Diftitox (18 μg/kg, i.v.) followed by several cell-lysate vaccinations. Host peripheral blood mononuclear cell lysates combined with CpG-ODN, and Montanide^® ISA 51 were locally applied. In vitro studies demonstrated that canine T_reg are a target of Denileukin Diftitox. The suppression of T-cell proliferation by T_reg was abolished by addition of Denileukin Diftitox (10 nM). An increase of proliferation of median 300% (range: 200%–425%) was observed. No change in donor chimerism was observed after administration of Denileukin Diftitox and vaccination. This study highlights that application of Denileukin Diftitox resulted in a depletion of T_reg followed by an increase of immune response in vitro. This effect could not be confirmed in vivo even if the immune system was stimulated by vaccinations.     

Inverse relationship between heat stable enterotoxin-b induced fluid accumulation and adherence of F4ac-positive enterotoxigenic Escherichia coli in ligated jejunal loops of F4ab/ac fimbria receptor-positive swine

Heat-labile enterotoxin (LT) produced by enterotoxigenic Escherichia coli (ETEC) increases bacterial adherence to porcine enterocytes in vitro and enhances small intestinal colonization in swine. Heat-stable enterotoxin-b (STb) is not known to affect colonization; however, through an induction of net fluid accumulation it might reduce bacterial adherence. The relationship between fluid accumulation and bacterial adherence in jejunal loops inoculated with ETEC strains that produce LT, STb, both, or neither toxin was studied. Ligated jejunal loops were constructed in weaned Yorkshire pigs in two independent experiments (Exp. 1, n = 5, 8-week-old; Exp. 2, n = 6, 6–8-week-old). Each pig was inoculated with six F4ac⁺ E. coli strains: (1) LT⁺, STb⁺ parent (WAM2317); (2) STb^? (ΔestB) mutant (MUN297); (3) MUN297 complemented with STb (MUN298); (4) LT^? STb^? (ΔeltAB ΔestB) mutant (MUN300); (5) MUN300 complemented with LT (MUN301); and (6) 1836-2 (non-enterotoxigenic, wild-type). Pigs were confirmed to be K88 (F4)ab/ac receptor-positive in Exp. 2 by testing for intestinal mucin-type glycoproteins and inferred to be receptor-positive in both Exp. 1 and 2 based on histopathologic evidence of bacterial adherence. Strains that produced STb induced marked fluid accumulation with the response (ml/cm) to WAM2317 and MUN298 significantly greater than that to the other strains (P < 0.0001). Conversely, bacterial adherence scores based on immunohistochemistry and CFU/g of washed mucosa were both lowest in the strains that expressed STb and highest in those that did not. For the two experiments combined, the Pearson correlation coefficient (R) between fluid volume (ml/cm) and log CFU per gram was ?0.57021 (P < 0.0001); R² = 0.3521 (n = 197). These results support the hypothesis that enterotoxin-induced fluid accumulation flushes progeny organisms into the lumen of the bowel, thereby increasing the likelihood of fecal shedding and transmission of the pathogen to new hosts.     

Leukocytic responses and intestinal mucin dynamics of broilers protected with Enterococcus faecium EF55 and challenged with Salmonella Enteritidis

The protective effect of Enterococcus faecium EF55 in chickens challenged with Salmonella enterica serovar Enteritidis phage type 4 (SE PT4) was assessed. The antibacterial effect on the bacterial microflora in the small intestine in relation to white blood cell count, phenotyping of peripheral blood and intestinal lymphocytes, functional activity of lymphocytes and phagocytes and mucin quantitation were investigated. Day-old chicks (85) were randomly divided into four groups. The probiotic group (EF) and Salmonella + probiotic group (EFSE) received E. faecium EF55 (10⁹ CFU – 3 g/group/day) for 21 days. The Salmonella group (SE) and EFSE group were infected with Salmonella Enteritidis (10⁸ CFU in 0.2 ml PBS) in a single dose per os on day four of the experiment. The control group chicks (C) were fed a commercial diet without added bacteria. Supplementation of EF55 in the diet of the chickens in the EFSE group, challenged with S. Enteritidis, caused the density of the intestinal mucin layer to increase significantly in non-specific regions (duodenum and jejunum), but decrease significantly in target regions (caeca) for S. Enteritidis. Probiotic treatment also appeared to result in a significantly higher number of lymphocytes in peripheral blood and a tendency to increase CD3, CD4, CD8, and IgM positive cells 3 days post-infection with S. Enteritidis. The results demonstrated an antibacterial effect and suggested that EF55 had a moderating effect on intestinal mucin production and leukocytic response in the early phase of S. Enteritidis infection.     

Feeding Redberry Juniper (Juniperus pinchotii) at Weaning Increases Juniper Consumption by Goats on Pasture

Redberry (Juniperus pinchotii Sudw.) and ashe (Juniperus ashei Buchh.) juniper dominate rangelands throughout central Texas. Our objective was to attempt to improve the efficacy of goats as a biological control mechanism for juniper through behavioral training. Conditioning sheep and goats to increase the palatability of chemically defended plants can be a useful tool in brush control. Previous research illustrated that goats can be conditioned to consume more juniper while in individual pens when foraging choices are limited. To test whether this creates a longer-lasting increase in juniper preference, we determined if goats would continue to consume juniper on pasture for one year after being fed juniper in individual pens for 14 d. Female Boer-cross goats (n = 40) were randomly divided into two treatments: conditioned and naive to juniper. At approximately 12 mo of age, conditioned goats were placed in individual pens and fed redberry juniper 1 h daily for 14 d, while naive goats received only alfalfa pellets to meet maintenance requirements. After the pen-feeding phase of the study, goats were placed in one of four pastures (10 goats · pasture^?1) for 12 mo. Two pastures housed conditioned goats, and two pastures housed naive goats at a moderate stocking rate (1 animal unit · yr^?1 · 8 ha^?1). Bite count surveys were conducted twice per month, while herbaceous standing crop and monoterpene levels were measured once per month. Juniper preference varied monthly; however, conditioned goats consistently ate more (P < 0.05) juniper than naive goats except for April, when the study began, and March, when the study ended. When selection of herbaceous forages decreased, conditioned goats increased selection of juniper, while naive goats increased selection of other palatable shrubs. Seasonal changes of monoterpene levels in juniper had no apparent effect on juniper preference. We contend that feeding juniper at weaning will increase use of the plant in grazing situations.     

Supplementing Guinea grass with fresh sweet potato foliage for milk production by Bunaji and N'Dama cows in early lactation

Forage from three sweet potato cultivars (A = TIS-87/0087; B = TIS-8164; C = TIS-2532.OP.1.13 at 30% daily dry matter intake), dried brewers' grains (DBG) and cottonseed meal (CSM) each at 2.5 kg were supplemented to Guinea grass (GG) to form four diets: Diet A = GG + TIS-87/0087; Diet B = GG + TIS-8164; Diet C = GG + TIS-2532.OP.1.13, and Diet D = GG + DBG + CSM (as control). Treatments were assigned as 4 × 4 Latin squares design over 60 days (10-day adaptation and 5-day sampling) using Bunaji and N'Dama cows in early lactation. The 48-h rumen dry matter (DM) degradation ranged (P < 0.01) from 407 g kg^? 1 DM for GG to 791 g kg^? 1 DM for sweet potato cultivar TIS-87/0087. Bunaji dry matter intake varied (P < 0.05) between 7.1 kg day^? 1 in Diet B and 8.9 kg day^? 1 in Diet D, but was similar (P > 0.05) among diets for the N'Dama cows. The metabolisable energy (ME) intakes were higher for Diet D although, it recorded the least efficiency of ME utilization for milk production. Milk yields were significantly (P < 0.01) higher in the Bunaji than the N'Dama cows, which is typical of their true breed differences. Total solids, ash, protein, fat, and sugar contents of the milk were similar among diets for both cow breeds, except Bunaji ash contents that ranged (P < 0.05) from 0.77 g 100 g^? 1 for Diet B to 0.83 g 100 g^? 1 for Diet D. The results suggest that sweet potato forage could be utilized as whole or partial replacement for DBG and CSM to save cost under smallholder farming systems.     

Effect of molecular weight and concentration of legume condensed tannins on in vitro larval migration inhibition of Haemonchus contortus

The effect of molecular weight of condensed tannins (CT) from a variety of warm-season perennial legumes commonly consumed by sheep and goats on anthelmintic activity has not been previously explored. The objectives of this study were to determine if molecular weight of CT from warm-season perennial legumes could predict the biological activity of CT relative to anthelmintic activity against ivermectin resistant L3 stage Haemonchus contortus (HC) using a larval migration inhibition (LMI) assay. A second objective was to determine if CT from warm-season perennial legumes possess anthelmintic properties against L3 stage (HC). Lespedeza stuevei had the greatest concentration of total condensed tannin (TCT; 11.7%), whereas, with the exception of Arachis glabrata, a CT-free negative control, Leucaena retusa had the least TCT (3.3%). Weight-average molecular weight of CT ranged from 552 Da for L. stuevei to 1483 Da for Lespedeza cuneata. The treatments demonstrating the greatest percent LMI were L. retusa, L. stuevei and Acacia angustissima var. hirta (65.4%, 63.1% and 42.2%, respectively). The ivermectin treatment had the smallest percent LMI (12.5%) against ivermectin resistant L3 HC. There was a weak correlation (R² = 0.34; P = 0.05) between CT M_W and percent LMI, suggesting that molecular weight of CT is a weak contributing factor to CT biological activity as it relates to LMI of L3 stage HC. L. stuevei, L. retusa and A. angustissima var. hirta STP5 warrant further evaluation of anthelmintic properties in vivo.     

Expansion of intracellular IFN-γ positive lymphocytes during Mycoplasma agalactiae infection in sheep

A method to assess the expansion of antigen-specific intracellular IFN-γ positive T cell subsets during the infection will be helpful for a better understanding of mycoplasmal infections physiopathology in the sheep. We analysed the percentage of antigen-specific lymphocytes positive for intracellular IFN-γ during the infection of sheep with Mycoplasma agalactiae by culturing peripheral blood mononuclear cells of infected or uninfected animals with irradiated M. agalactiae. The expansion of antigen-specific IFN-γ positive lymphocytes in infected sheep was initially sustained by CD4⁺ T cells at day 15 after infection, when antigen specific IgG start to be detectable, followed by CD8/IFN-γ double positive cells. γδ T-cells were not expanded at any time point analysed. IFNγ⁺ T cells disappear 60 days after infection, suggesting that antigen specific IFNγ⁺ T cells, mainly detected in the early phase of the disease, could be useful to understand the role of cell-mediated immunity during M. agalactiae infection.     

Pharmacokinetic,metabolism and withdrawal time of orphenadrine in camels (Camelus dromedarius) after intravenous administration

The pharmacokinetics of orphenadrine (ORPH) following a single intravenous (i.v.) dose was investigated in six camels (Camelus dormedarius). Orphenadrine was extracted from the plasma using a simple sensitive liquid–liquid extraction method and determined by gas chromatography/mass spectrometry (GC/MS). Following i.v. administration plasma concentrations of ORPH decline bi-exponentially with distribution half-life (t_1/2α) of 0.50 ± 0.07 h, elimination half-life (t_1/2β) of 3.57 ± 0.55 h, area under the time concentration curve (AUC) of 1.03 ± 0.10 g/h l⁻¹. The volume of distribution at steady state (Vd_ss) 1.92 ± 0.22 l kg⁻¹, volume of the central compartment of the two compartment pharmacokinetic model (V_c) 0.87 ± 0.09 l kg⁻¹, and total body clearance (Cl_T) of 0.60 ± 0.09 l/h kg⁻¹. Three orphenadrine metabolites were identified in urine samples of camels. The first metabolite N-desmethyl-orphenadrine resulted from N-dealkylation of ORPH with molecular ion m/z 255. The second N,N-didesmethyl-orphenadrine, resulted from N-didesmethylation with molecular ion m/z 241. The third metabolite, hydroxyl-orphenadrine, resulted from the hydroxylation of ORPH with molecular ion m/z 285. ORPH and its metabolites in camel were extensively eliminated in conjugated form. ORPH remains detectable in camel urine for three days after i.v. administration of a single dose of 350 mg orphenadrine aspartate.     

A comparison of three different methodologies for evaluating Rhipicephalus (Boophilus) microplus susceptibility to topical spray compounds

This study aimed to compare three different methodologies (Adult Immersion Tests, field trials with naturally infected animals, and a Stall Test using artificially infested cattle) to evaluate the efficacy of two topical formulations that we administered as whole body sprays (15% Cypermethrin + 30% Chlorpyriphos + 15% Fenthion–Colosso^® FC 30, Ouro Fino Agronegócios; and 60% Dichlorvos + 20% Chlorpyriphos–Ectofós^®, Vallée Saúde Animal Ltd.), against a susceptible strain of Rhipicephalus (Boophilus) microplus. To achieve this objective, two natural infestation trials were conducted, as well as two artificial infestation trials (Stall Tests) and two Adult Immersion Tests (AIT). The AIT results showed that both spray formulations achieved 100% efficacy against R. (B.) microplus fully engorged females. However, when observing results obtained by field trials (natural infestations) and Stall Tests, none of these topically applied compounds reached 100% efficacy or affected the reproductive capacity of the fully engorged female ticks. Additional studies must be conducted to compare these in vivo methodologies with different in vitro techniques, such as the Larval Packet Test. However, based on results obtained here, we can conclude that depending on the spray formulations used, the AIT can overestimate acaricidal efficacy and values of reproductive efficiency of such compounds against R (B.) microplus. Specifically, when dealing with spray formulations in the Stall Tests, the period of residual action can increase because these animals are sheltered from contact with environmental factors that might interfere with the efficacy of the products tested. It may be necessary to take in vivo trial results into consideration (such as field trials with naturally infested animals or Stall Tests) to standardize a specific in vitro assay, such as the Adult Immersion Test.